doc_id	sent_index	relation_id	relation	trigger	trigger_offset	arg_num	arg_head	arg_protein	arg_domain	arg_site	arg_sugar	arg_head_offset	arg_base_np	arg_base_np_offset	arg_np	arg_np_offset	sent_text
21389326	0	0	theme	complex	88:94	arg1	assembly					43:50	the extracellular assembly	25:50	the extracellular assembly of the hematopoietic Flt3 signaling complex	25:94	Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex.
21389326	1	1	theme	signaling	292:300	arg1	cascades					302:309	signaling cascades	292:309	signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells	292:421	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	2	2	theme	acute	586:590	arg1	leukemia					600:607	acute myeloid leukemia	586:607	acute myeloid leukemia	586:607	However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia.
21389326	4	3	theme	extracellular	889:901	arg1	domain					903:908	extracellular domain 3	889:910	extracellular domain 3 of Flt3	889:918	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	4	3	theme	extracellular	889:901	arg1	Flt3					915:918	Flt3	915:918	Flt3	915:918	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	1	4	theme	Fms-like	145:152	arg1	receptor					170:177	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3	97:179	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3)	97:186	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	1	4	theme	Fms-like	145:152	arg1	Flt3					182:185	Flt3	182:185	Flt3	182:185	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	7	5	theme	FL	1442:1443	arg1	reminiscent					1445:1455	FL reminiscent	1442:1455	FL reminiscent of a "lock-and-key" binding mode	1442:1488	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
21389326	0	6	theme	signaling	78:86	arg1	complex					88:94	the hematopoietic Flt3 signaling complex	55:94	the hematopoietic Flt3 signaling complex	55:94	Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex.
21389326	3	7	theme	Flt3	655:658	arg1	complex					676:682	the Flt3 ligand-receptor complex	651:682	the Flt3 ligand-receptor complex	651:682	Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date.
21389326	3	8	theme	structural	630:639	arg1	basis					641:645	the structural basis	626:645	the structural basis for the Flt3 ligand-receptor complex	626:682	Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date.
21389326	1	9	theme	tyrosine	154:161	arg1	receptor					170:177	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3	97:179	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3)	97:186	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	1	9	theme	tyrosine	154:161	arg1	Flt3					182:185	Flt3	182:185	Flt3	182:185	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	1	10	theme	crucial	311:317	arg1	cascades					302:309	signaling cascades	292:309	signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells	292:421	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	6	11	theme	points	1227:1232	arg1	dissection					1195:1204	thermodynamic dissection	1181:1204	thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event	1181:1283	Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty.
21389326	7	12	theme	preformed	1413:1421	arg1	epitope					1431:1437	a single preformed binding epitope	1404:1437	a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode	1404:1488	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
21389326	1	13	theme	central	222:228	arg1	roles					230:234	central roles	222:234	central roles	222:234	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	2	14	theme	hematologic	494:504	arg1	malignancies					506:517	hematologic malignancies	494:517	hematologic malignancies	494:517	However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia.
21389326	3	15	theme	unanticipated	698:710	arg1	assembly					726:733	an unanticipated extracellular assembly	695:733	an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date	695:789	Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date.
21389326	2	16	theme	clinical	566:573	arg1	target					575:580	clinical target	566:580	clinical target for acute myeloid leukemia	566:607	However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia.
21389326	7	17	theme	binding	1423:1429	arg1	epitope					1431:1437	a single preformed binding epitope	1404:1437	a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode	1404:1488	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
21389326	4	18	theme	homotypic	964:972	arg1	interactions					983:994	homotypic receptor interactions	964:994	homotypic receptor interactions	964:994	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	4	19	theme	interactions	983:994	arg1	devoid					954:959	devoid	954:959	devoid	954:959	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	5	20	with	Comparisons	997:1007	arg1	receptors					1033:1041	homologous receptors	1022:1041	homologous receptors	1022:1041	Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly.
21389326	3	21	theme	extracellular	712:724	arg1	assembly					726:733	an unanticipated extracellular assembly	695:733	an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date	695:789	Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date.
21389326	6	22	theme	binding	1271:1277	arg1	event					1279:1283	a pronounced enthalpically driven binding event	1237:1283	a pronounced enthalpically driven binding event	1237:1283	Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty.
21389326	7	23	theme	FL	1372:1373	arg1	complex					1375:1381	the high-affinity Flt3:FL complex	1349:1381	the high-affinity Flt3:FL complex	1349:1381	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
21389326	1	24	theme	kinase	163:168	arg1	receptor					170:177	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3	97:179	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3)	97:186	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	1	24	theme	kinase	163:168	arg1	Flt3					182:185	Flt3	182:185	Flt3	182:185	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	2	25	theme	receptors	481:489	arg1	one					446:448	one	446:448	one	446:448	However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia.
21389326	2	25	theme	receptors	481:489	arg1	receptors					481:489	the most frequently mutated receptors	453:489	the most frequently mutated receptors in hematologic malignancies	453:517	However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia.
21389326	6	26	theme	driven	1264:1269	arg1	event					1279:1283	a pronounced enthalpically driven binding event	1237:1283	a pronounced enthalpically driven binding event	1237:1283	Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty.
21389326	7	27	theme	single	1406:1411	arg1	epitope					1431:1437	a single preformed binding epitope	1404:1437	a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode	1404:1488	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
21389326	7	28	theme	"	1475:1475	arg1	mode					1485:1488	a "lock-and-key" binding mode	1460:1488	a "lock-and-key" binding mode	1460:1488	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
21389326	4	29	theme	ternary	938:944	arg1	complex					946:952	a ternary complex	936:952	a ternary complex devoid of homotypic receptor interactions	936:994	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	7	30	from	epitope	1431:1437	arg1	reminiscent					1445:1455	FL reminiscent	1442:1455	FL reminiscent of a "lock-and-key" binding mode	1442:1488	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
21389326	4	31	theme	devoid	954:959	arg1	complex					946:952	a ternary complex	936:952	a ternary complex devoid of homotypic receptor interactions	936:994	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	4	32	theme	binding	849:855	arg1	epitope					857:863	a remarkably compact binding epitope	828:863	a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3	828:918	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	2	33	theme	mutated	473:479	arg1	receptors					481:489	the most frequently mutated receptors	453:489	the most frequently mutated receptors in hematologic malignancies	453:517	However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia.
21389326	6	34	theme	pronounced	1239:1248	arg1	event					1279:1283	a pronounced enthalpically driven binding event	1237:1283	a pronounced enthalpically driven binding event	1237:1283	Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty.
21389326	0	35	theme	Structural	0:9	arg1	insights					11:18	Structural insights	0:18	Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex	0:94	Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex.
21389326	5	36	theme	assembly	1158:1165	arg1	features					1123:1130	the unique features	1112:1130	the unique features of the Flt3 extracellular assembly	1112:1165	Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly.
21389326	2	37	theme	myeloid	592:598	arg1	leukemia					600:607	acute myeloid leukemia	586:607	acute myeloid leukemia	586:607	However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia.
21389326	7	38	theme	lock-and-key	1463:1474	arg1	mode					1485:1488	a "lock-and-key" binding mode	1460:1488	a "lock-and-key" binding mode	1460:1488	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
21389326	7	39	theme	mode	1485:1488	arg1	reminiscent					1445:1455	FL reminiscent	1442:1455	FL reminiscent of a "lock-and-key" binding mode	1442:1488	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
21389326	5	40	theme	unique	1116:1121	arg1	features					1123:1130	the unique features	1112:1130	the unique features of the Flt3 extracellular assembly	1112:1165	Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly.
21389326	3	41	theme	other	746:750	arg1	complex					761:767	any other RTKIII/V complex	742:767	any other RTKIII/V complex characterized to date	742:789	Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date.
21389326	4	42	theme	Flt3	819:822	arg1	dimerization					803:814	dimerization	803:814	dimerization of Flt3	803:822	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	1	43	theme	class	101:105	arg1	receptor					170:177	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3	97:179	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3)	97:186	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	1	43	theme	class	101:105	arg1	Flt3					182:185	Flt3	182:185	Flt3	182:185	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	5	44	theme	available	1047:1055	arg1	data					1069:1072	available mutagenesis data	1047:1072	available mutagenesis data for FL	1047:1079	Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly.
21389326	5	45	theme	mutagenesis	1057:1067	arg1	data					1069:1072	available mutagenesis data	1047:1072	available mutagenesis data for FL	1047:1079	Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly.
21389326	4	46	theme	receptor	974:981	arg1	interactions					983:994	homotypic receptor interactions	964:994	homotypic receptor interactions	964:994	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	5	47	with	data	1069:1072	arg1	receptors					1033:1041	homologous receptors	1022:1041	homologous receptors	1022:1041	Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly.
21389326	0	48	theme	extracellular	29:41	arg1	assembly					43:50	the extracellular assembly	25:50	the extracellular assembly of the hematopoietic Flt3 signaling complex	25:94	Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex.
21389326	1	49	theme	immune	261:266	arg1	system					268:273	the immune system	257:273	the immune system	257:273	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	4	50	theme	Flt3	915:918	arg1	domain					903:908	extracellular domain 3	889:910	extracellular domain 3 of Flt3	889:918	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	4	50	theme	Flt3	915:918	arg1	Flt3					915:918	Flt3	915:918	Flt3	915:918	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	1	51	theme	hematopoietic	358:370	arg1	progenitors					372:382	hematopoietic progenitors	358:382	hematopoietic progenitors	358:382	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	5	52	theme	Flt3	1139:1142	arg1	assembly					1158:1165	the Flt3 extracellular assembly	1135:1165	the Flt3 extracellular assembly	1135:1165	Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly.
21389326	6	53	theme	entropic	1299:1306	arg1	penalty					1308:1314	an entropic penalty	1296:1314	an entropic penalty	1296:1314	Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty.
21389326	1	54	theme	progenitors	372:382	arg1	homeostasis					343:353	homeostasis	343:353	homeostasis	343:353	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	1	54	theme	progenitors	372:382	arg1	development					327:337	the development	323:337	the development	323:337	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	5	55	theme	Flt3	1012:1015	arg1	data					1069:1072	available mutagenesis data	1047:1072	available mutagenesis data for FL	1047:1079	Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly.
21389326	5	55	theme	Flt3	1012:1015	arg1	Comparisons					997:1007	Comparisons	997:1007	Comparisons of Flt3 with homologous receptors	997:1041	Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly.
21389326	4	56	theme	domain	903:908	arg1	tip					882:884	the tip	878:884	the tip of extracellular domain 3 of Flt3	878:918	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	1	57	theme	receptor	111:118	arg1	receptor					170:177	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3	97:179	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3)	97:186	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	1	57	theme	receptor	111:118	arg1	Flt3					182:185	Flt3	182:185	Flt3	182:185	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	3	58	theme	ligand-receptor	660:674	arg1	complex					676:682	the Flt3 ligand-receptor complex	651:682	the Flt3 ligand-receptor complex	651:682	Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date.
21389326	3	59	theme	RTKIII/V	752:759	arg1	complex					761:767	any other RTKIII/V complex	742:767	any other RTKIII/V complex characterized to date	742:789	Here, we report the structural basis for the Flt3 ligand-receptor complex and unveil an unanticipated extracellular assembly unlike any other RTKIII/V complex characterized to date.
21389326	5	60	theme	homologous	1022:1031	arg1	receptors					1033:1041	homologous receptors	1022:1041	homologous receptors	1022:1041	Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly.
21389326	2	61	from	receptors	481:489	arg1	malignancies					506:517	hematologic malignancies	494:517	hematologic malignancies	494:517	However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia.
21389326	1	62	theme	tyrosine	120:127	arg1	receptor					170:177	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3	97:179	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3)	97:186	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	1	62	theme	tyrosine	120:127	arg1	Flt3					182:185	Flt3	182:185	Flt3	182:185	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	6	63	theme	formation	1217:1225	arg1	points					1227:1232	complex formation points	1209:1232	complex formation points	1209:1232	Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty.
21389326	5	64	theme	extracellular	1144:1156	arg1	assembly					1158:1165	the Flt3 extracellular assembly	1135:1165	the Flt3 extracellular assembly	1135:1165	Comparisons of Flt3 with homologous receptors and available mutagenesis data for FL have allowed us to rationalize the unique features of the Flt3 extracellular assembly.
21389326	2	65	theme	prognostic	544:553	arg1	factor					555:560	a major prognostic factor	536:560	a major prognostic factor	536:560	However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia.
21389326	1	66	theme	antigen-presenting	388:405	arg1	cells					417:421	antigen-presenting dendritic cells	388:421	antigen-presenting dendritic cells	388:421	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	6	67	theme	complex	1209:1215	arg1	points					1227:1232	complex formation points	1209:1232	complex formation points	1209:1232	Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty.
21389326	1	68	theme	kinase	129:134	arg1	receptor					170:177	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3	97:179	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3)	97:186	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	1	68	theme	kinase	129:134	arg1	Flt3					182:185	Flt3	182:185	Flt3	182:185	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	4	69	located	localized	865:873	arg1	tip					882:884	the tip	878:884	the tip of extracellular domain 3 of Flt3	878:918	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	4	69	located	localized	865:873	arg2	epitope					857:863	a remarkably compact binding epitope	828:863	a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3	828:918	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	1	70	theme	cytokine	196:203	arg1	FL					213:214	FL	213:214	FL	213:214	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	1	70	theme	cytokine	196:203	arg1	ligand					205:210	its cytokine ligand	192:210	its cytokine ligand (FL)	192:215	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	2	71	theme	major	538:542	arg1	factor					555:560	a major prognostic factor	536:560	a major prognostic factor	536:560	However, Flt3 is also one of the most frequently mutated receptors in hematologic malignancies and is currently a major prognostic factor and clinical target for acute myeloid leukemia.
21389326	1	72	theme	dendritic	407:415	arg1	cells					417:421	antigen-presenting dendritic cells	388:421	antigen-presenting dendritic cells	388:421	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	7	73	theme	Flt3	1367:1370	arg1	complex					1375:1381	the high-affinity Flt3:FL complex	1349:1381	the high-affinity Flt3:FL complex	1349:1381	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
21389326	0	74	theme	Flt3	73:76	arg1	complex					88:94	the hematopoietic Flt3 signaling complex	55:94	the hematopoietic Flt3 signaling complex	55:94	Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex.
21389326	1	75	theme	cells	417:421	arg1	homeostasis					343:353	homeostasis	343:353	homeostasis	343:353	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	1	75	theme	cells	417:421	arg1	development					327:337	the development	323:337	the development	323:337	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	7	76	theme	antagonist	1521:1530	arg1	design					1532:1537	antagonist design	1521:1537	antagonist design	1521:1537	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
21389326	1	77	theme	RTKIII	137:142	arg1	receptor					170:177	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3	97:179	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3)	97:186	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	1	77	theme	RTKIII	137:142	arg1	Flt3					182:185	Flt3	182:185	Flt3	182:185	The class III receptor tyrosine kinase (RTKIII) Fms-like tyrosine kinase receptor 3 (Flt3) and its cytokine ligand (FL) play central roles in hematopoiesis and the immune system, by establishing signaling cascades crucial for the development and homeostasis of hematopoietic progenitors and antigen-presenting dendritic cells.
21389326	0	78	theme	hematopoietic	59:71	arg1	complex					88:94	the hematopoietic Flt3 signaling complex	55:94	the hematopoietic Flt3 signaling complex	55:94	Structural insights into the extracellular assembly of the hematopoietic Flt3 signaling complex.
21389326	7	79	theme	high-affinity	1353:1365	arg1	complex					1375:1381	the high-affinity Flt3:FL complex	1349:1381	the high-affinity Flt3:FL complex	1349:1381	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
21389326	6	80	theme	thermodynamic	1181:1193	arg1	dissection					1195:1204	thermodynamic dissection	1181:1204	thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event	1181:1283	Furthermore, thermodynamic dissection of complex formation points to a pronounced enthalpically driven binding event coupled to an entropic penalty.
21389326	4	81	theme	compact	841:847	arg1	epitope					857:863	a remarkably compact binding epitope	828:863	a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3	828:918	FL induces dimerization of Flt3 via a remarkably compact binding epitope localized at the tip of extracellular domain 3 of Flt3, and it invokes a ternary complex devoid of homotypic receptor interactions.
21389326	7	82	theme	binding	1477:1483	arg1	mode					1485:1488	a "lock-and-key" binding mode	1460:1488	a "lock-and-key" binding mode	1460:1488	Together, our data suggest that the high-affinity Flt3:FL complex is driven in part by a single preformed binding epitope on FL reminiscent of a "lock-and-key" binding mode, thereby setting the stage for antagonist design.
19202066	0	0	theme	glucosamine	82:92	arg1	modification					94:105	O-linked beta-N-acetyl glucosamine modification	59:105	O-linked beta-N-acetyl glucosamine modification	59:105	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.
19202066	8	1	from	IKK	1523:1525	arg1	oncogenesis					1530:1540	oncogenesis	1530:1540	oncogenesis	1530:1540	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.
19202066	2	2	theme	positive	322:329	arg1	loop					340:343	the positive feedback loop	318:343	the positive feedback loop between IKK-NF-kappaB and glycolysis	318:380	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.
19202066	0	3	theme	beta-N-acetyl	68:80	arg1	modification					94:105	O-linked beta-N-acetyl glucosamine modification	59:105	O-linked beta-N-acetyl glucosamine modification	59:105	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.
19202066	3	4	theme	human	663:667	arg1	fibroblasts					669:679	transformed human fibroblasts	651:679	transformed human fibroblasts	651:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	7	5	theme	IKKbeta	1152:1158	arg1	O-GlcNAcylation					1133:1147	O-GlcNAcylation	1133:1147	O-GlcNAcylation of IKKbeta	1133:1158	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.
19202066	5	6	theme	IKKbeta	902:908	arg1	O-GlcNAcylation					883:897	the O-GlcNAcylation	879:897	the O-GlcNAcylation of IKKbeta	879:908	We also found that high glucose induced the O-GlcNAcylation of IKKbeta and sustained the TNFalpha-dependent IKKbeta activity.
19202066	8	7	theme	activity	1420:1427	arg1	enhancement					1395:1405	the enhancement	1391:1405	the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis	1391:1540	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.
19202066	6	8	theme	O-GlcNAcase	979:989	arg1	streptozotocin					1001:1014	the O-GlcNAcase inhibitor streptozotocin	975:1014	the O-GlcNAcase inhibitor streptozotocin	975:1014	Moreover, the O-GlcNAcase inhibitor streptozotocin intensified O-GlcNAcylation and concomitant activating phosphorylation of IKKbeta.
19202066	8	9	theme	NF-kappaB	1410:1418	arg1	activity					1420:1427	NF-kappaB activity	1410:1427	NF-kappaB activity	1410:1427	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.
19202066	7	10	theme	catalytic	1322:1330	arg1	activity					1332:1339	its catalytic activity	1318:1339	its catalytic activity	1318:1339	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.
19202066	4	11	theme	activating	741:750	arg1	phosphorylation					752:766	the activating phosphorylation	737:766	the activating phosphorylation of IKK	737:773	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.
19202066	8	12	theme	novel	1371:1375	arg1	mechanism					1377:1385	a novel mechanism	1369:1385	a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis	1369:1540	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.
19202066	7	13	theme	phosphorylation	1246:1260	arg1	domain					1198:1203	the C-terminal domain	1183:1203	the C-terminal domain	1183:1203	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.
19202066	7	13	theme	phosphorylation	1246:1260	arg1	site					1262:1265	an inactivating phosphorylation site	1230:1265	an inactivating phosphorylation site	1230:1265	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.
19202066	4	14	theme	O-GlcNAcylated	710:723	arg1	IKKbeta					725:731	the O-GlcNAcylated IKKbeta	706:731	the O-GlcNAcylated IKKbeta	706:731	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.
19202066	2	15	theme	IKK-NF-kappaB	263:275	arg1	pathway					277:283	the IKK-NF-kappaB pathway	259:283	the IKK-NF-kappaB pathway	259:283	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.
19202066	1	16	theme	critical	159:166	arg1	role					168:171	a critical role	157:171	a critical role	157:171	The IkappaB kinase (IKK)-NF-kappaB pathway plays a critical role in oncogenesis.
19202066	3	17	theme	complex	502:508	arg1	IKKbeta					470:476	IKKbeta	470:476	IKKbeta	470:476	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	17	theme	complex	502:508	arg1	component					481:489	a component	479:489	a component of the IKK complex	479:508	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	8	18	theme	positive	1458:1465	arg1	regulation					1476:1485	positive feedback regulation	1458:1485	positive feedback regulation	1458:1485	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.
19202066	5	19	theme	high	858:861	arg1	glucose					863:869	high glucose	858:869	high glucose	858:869	We also found that high glucose induced the O-GlcNAcylation of IKKbeta and sustained the TNFalpha-dependent IKKbeta activity.
19202066	3	20	link	O-linked	544:551	arg1	O-GlcNAc					580:587	O-GlcNAc	580:587	O-GlcNAc	580:587	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	20	link	O-linked	544:551	arg1	glucosamine					567:577	O-linked beta-N-acetyl glucosamine	544:577	O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts	544:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	7	21	theme	IKKbeta	1284:1290	arg1	O-GlcNAcylation					1292:1306	IKKbeta O-GlcNAcylation	1284:1306	IKKbeta O-GlcNAcylation	1284:1306	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.
19202066	5	22	theme	IKKbeta	947:953	arg1	activity					955:962	the TNFalpha-dependent IKKbeta activity	924:962	the TNFalpha-dependent IKKbeta activity	924:962	We also found that high glucose induced the O-GlcNAcylation of IKKbeta and sustained the TNFalpha-dependent IKKbeta activity.
19202066	2	23	theme	cell	423:426	arg1	transformation					428:441	oncogene-induced cell transformation	406:441	oncogene-induced cell transformation	406:441	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.
19202066	6	24	theme	inhibitor	991:999	arg1	streptozotocin					1001:1014	the O-GlcNAcase inhibitor streptozotocin	975:1014	the O-GlcNAcase inhibitor streptozotocin	975:1014	Moreover, the O-GlcNAcase inhibitor streptozotocin intensified O-GlcNAcylation and concomitant activating phosphorylation of IKKbeta.
19202066	3	25	theme	O-linked	544:551	arg1	O-GlcNAc					580:587	O-GlcNAc	580:587	O-GlcNAc	580:587	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	25	theme	O-linked	544:551	arg1	glucosamine					567:577	O-linked beta-N-acetyl glucosamine	544:577	O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts	544:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	7	26	theme	C-terminal	1187:1196	arg1	domain					1198:1203	the C-terminal domain	1183:1203	the C-terminal domain	1183:1203	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.
19202066	7	26	theme	C-terminal	1187:1196	arg1	site					1262:1265	an inactivating phosphorylation site	1230:1265	an inactivating phosphorylation site	1230:1265	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.
19202066	2	27	theme	oncogene-induced	406:421	arg1	transformation					428:441	oncogene-induced cell transformation	406:441	oncogene-induced cell transformation	406:441	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.
19202066	4	28	theme	p65/NF-kappaB	793:805	arg1	knockdown					807:815	p65/NF-kappaB knockdown	793:815	p65/NF-kappaB knockdown	793:815	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.
19202066	8	29	theme	feedback	1467:1474	arg1	regulation					1476:1485	positive feedback regulation	1458:1485	positive feedback regulation	1458:1485	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.
19202066	3	30	from	glucosamine	567:577	arg1	fibroblasts					628:638	p53-deficient mouse embryonic fibroblasts	598:638	p53-deficient mouse embryonic fibroblasts (MEFs)	598:645	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	30	from	glucosamine	567:577	arg1	fibroblasts					669:679	transformed human fibroblasts	651:679	transformed human fibroblasts	651:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	30	from	glucosamine	567:577	arg1	MEFs					641:644	MEFs	641:644	MEFs	641:644	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	0	31	theme	p53	8:10	arg1	Loss					0:3	Loss	0:3	Loss of p53	0:10	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.
19202066	3	32	theme	IKK	498:500	arg1	complex					502:508	the IKK complex	494:508	the IKK complex	494:508	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	1	33	theme	IkappaB	112:118	arg1	IKK					128:130	IKK	128:130	IKK	128:130	The IkappaB kinase (IKK)-NF-kappaB pathway plays a critical role in oncogenesis.
19202066	1	33	theme	IkappaB	112:118	arg1	kinase					120:125	IkappaB kinase	112:125	The IkappaB kinase (IKK)-NF-kappaB pathway	108:149	The IkappaB kinase (IKK)-NF-kappaB pathway plays a critical role in oncogenesis.
19202066	8	34	theme	p53	1440:1442	arg1	loss					1432:1435	loss	1432:1435	loss	1432:1435	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.
19202066	2	35	theme	integral	389:396	arg1	role					398:401	an integral role	386:401	an integral role	386:401	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.
19202066	4	36	theme	p53-deficient	685:697	arg1	cells					699:703	p53-deficient cells	685:703	p53-deficient cells	685:703	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.
19202066	7	37	theme	inactivating	1233:1244	arg1	domain					1198:1203	the C-terminal domain	1183:1203	the C-terminal domain	1183:1203	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.
19202066	7	37	theme	inactivating	1233:1244	arg1	site					1262:1265	an inactivating phosphorylation site	1230:1265	an inactivating phosphorylation site	1230:1265	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.
19202066	3	38	mod	modified	530:537	arg1	IKKbeta					470:476	IKKbeta	470:476	IKKbeta	470:476	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	38	mod	modified	530:537	arg1	component					481:489	a component	479:489	a component of the IKK complex	479:508	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	38	mod	modified	530:537	arg3	glucosamine					567:577	O-linked beta-N-acetyl glucosamine	544:577	O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts	544:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	38	mod	modified	530:537	arg3	O-GlcNAc					580:587	O-GlcNAc	580:587	O-GlcNAc	580:587	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	0	39	theme	catalytic	21:29	arg1	activity					31:38	catalytic activity	21:38	catalytic activity of IKKbeta	21:49	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.
19202066	3	40	theme	mouse	612:616	arg1	fibroblasts					628:638	p53-deficient mouse embryonic fibroblasts	598:638	p53-deficient mouse embryonic fibroblasts (MEFs)	598:645	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	40	theme	mouse	612:616	arg1	MEFs					641:644	MEFs	641:644	MEFs	641:644	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	6	41	theme	IKKbeta	1090:1096	arg1	O-GlcNAcylation					1028:1042	O-GlcNAcylation	1028:1042	O-GlcNAcylation	1028:1042	Moreover, the O-GlcNAcase inhibitor streptozotocin intensified O-GlcNAcylation and concomitant activating phosphorylation of IKKbeta.
19202066	6	41	theme	IKKbeta	1090:1096	arg1	phosphorylation					1071:1085	concomitant activating phosphorylation	1048:1085	concomitant activating phosphorylation of IKKbeta	1048:1096	Moreover, the O-GlcNAcase inhibitor streptozotocin intensified O-GlcNAcylation and concomitant activating phosphorylation of IKKbeta.
19202066	0	42	theme	IKKbeta	43:49	arg1	activity					31:38	catalytic activity	21:38	catalytic activity of IKKbeta	21:49	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.
19202066	7	43	theme	Mutational	1099:1108	arg1	analysis					1110:1117	Mutational analysis	1099:1117	Mutational analysis	1099:1117	Mutational analysis revealed that O-GlcNAcylation of IKKbeta occurred at Ser 733 in the C-terminal domain, which was identified as an inactivating phosphorylation site, suggesting that IKKbeta O-GlcNAcylation regulates its catalytic activity.
19202066	3	44	theme	embryonic	618:626	arg1	fibroblasts					628:638	p53-deficient mouse embryonic fibroblasts	598:638	p53-deficient mouse embryonic fibroblasts (MEFs)	598:645	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	44	theme	embryonic	618:626	arg1	MEFs					641:644	MEFs	641:644	MEFs	641:644	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	45	theme	beta-N-acetyl	553:565	arg1	O-GlcNAc					580:587	O-GlcNAc	580:587	O-GlcNAc	580:587	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	45	theme	beta-N-acetyl	553:565	arg1	glucosamine					567:577	O-linked beta-N-acetyl glucosamine	544:577	O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts	544:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	1	46	theme	kinase	120:125	arg1	pathway					143:149	The IkappaB kinase (IKK)-NF-kappaB pathway	108:149	The IkappaB kinase (IKK)-NF-kappaB pathway	108:149	The IkappaB kinase (IKK)-NF-kappaB pathway plays a critical role in oncogenesis.
19202066	2	47	contain	has	382:384	arg2	role					398:401	an integral role	386:401	an integral role	386:401	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.
19202066	2	47	contain	has	382:384	arg1	loop					340:343	the positive feedback loop	318:343	the positive feedback loop between IKK-NF-kappaB and glycolysis	318:380	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.
19202066	4	48	theme	glucose	820:826	arg1	depletion					828:836	glucose depletion	820:836	glucose depletion	820:836	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.
19202066	0	49	link	O-linked	59:66	arg1	modification					94:105	O-linked beta-N-acetyl glucosamine modification	59:105	O-linked beta-N-acetyl glucosamine modification	59:105	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.
19202066	2	50	theme	p53	313:315	arg1	absence					302:308	the absence	298:308	the absence of p53	298:315	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.
19202066	3	51	theme	transformed	651:661	arg1	fibroblasts					669:679	transformed human fibroblasts	651:679	transformed human fibroblasts	651:679	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	8	52	theme	enhanced	1492:1499	arg1	metabolism					1509:1518	enhanced glucose metabolism	1492:1518	enhanced glucose metabolism	1492:1518	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.
19202066	4	53	theme	IKK	771:773	arg1	phosphorylation					752:766	the activating phosphorylation	737:766	the activating phosphorylation of IKK	737:773	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.
19202066	4	53	theme	IKK	771:773	arg1	IKKbeta					725:731	the O-GlcNAcylated IKKbeta	706:731	the O-GlcNAcylated IKKbeta	706:731	In p53-deficient cells, the O-GlcNAcylated IKKbeta and the activating phosphorylation of IKK were decreased by p65/NF-kappaB knockdown or glucose depletion.
19202066	3	54	theme	p53-deficient	598:610	arg1	fibroblasts					628:638	p53-deficient mouse embryonic fibroblasts	598:638	p53-deficient mouse embryonic fibroblasts (MEFs)	598:645	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	3	54	theme	p53-deficient	598:610	arg1	MEFs					641:644	MEFs	641:644	MEFs	641:644	Here, we demonstrate that IKKbeta, a component of the IKK complex, was constitutively modified with O-linked beta-N-acetyl glucosamine (O-GlcNAc) in both p53-deficient mouse embryonic fibroblasts (MEFs) and transformed human fibroblasts.
19202066	5	55	theme	TNFalpha-dependent	928:945	arg1	activity					955:962	the TNFalpha-dependent IKKbeta activity	924:962	the TNFalpha-dependent IKKbeta activity	924:962	We also found that high glucose induced the O-GlcNAcylation of IKKbeta and sustained the TNFalpha-dependent IKKbeta activity.
19202066	0	56	theme	O-linked	59:66	arg1	modification					94:105	O-linked beta-N-acetyl glucosamine modification	59:105	O-linked beta-N-acetyl glucosamine modification	59:105	Loss of p53 enhances catalytic activity of IKKbeta through O-linked beta-N-acetyl glucosamine modification.
19202066	2	57	theme	glucose	232:238	arg1	metabolism					240:249	glucose metabolism	232:249	glucose metabolism	232:249	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.
19202066	1	58	theme	-NF-kappaB	132:141	arg1	pathway					143:149	The IkappaB kinase (IKK)-NF-kappaB pathway	108:149	The IkappaB kinase (IKK)-NF-kappaB pathway	108:149	The IkappaB kinase (IKK)-NF-kappaB pathway plays a critical role in oncogenesis.
19202066	6	59	theme	activating	1060:1069	arg1	phosphorylation					1071:1085	concomitant activating phosphorylation	1048:1085	concomitant activating phosphorylation of IKKbeta	1048:1096	Moreover, the O-GlcNAcase inhibitor streptozotocin intensified O-GlcNAcylation and concomitant activating phosphorylation of IKKbeta.
19202066	8	60	theme	glucose	1501:1507	arg1	metabolism					1509:1518	enhanced glucose metabolism	1492:1518	enhanced glucose metabolism	1492:1518	Taken together, we propose a novel mechanism for the enhancement of NF-kappaB activity by loss of p53, which evokes positive feedback regulation from enhanced glucose metabolism to IKK in oncogenesis.
19202066	2	61	theme	feedback	331:338	arg1	loop					340:343	the positive feedback loop	318:343	the positive feedback loop between IKK-NF-kappaB and glycolysis	318:380	Recently, we have shown that p53 regulates glucose metabolism through the IKK-NF-kappaB pathway and that, in the absence of p53, the positive feedback loop between IKK-NF-kappaB and glycolysis has an integral role in oncogene-induced cell transformation.
19202066	6	62	theme	concomitant	1048:1058	arg1	phosphorylation					1071:1085	concomitant activating phosphorylation	1048:1085	concomitant activating phosphorylation of IKKbeta	1048:1096	Moreover, the O-GlcNAcase inhibitor streptozotocin intensified O-GlcNAcylation and concomitant activating phosphorylation of IKKbeta.
16474139	4	0	theme	O-linked	896:903	arg1	oligosaccharides					905:920	O-linked oligosaccharides	896:920	O-linked oligosaccharides	896:920	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.
16474139	3	1	theme	coronavirus	480:490	arg1	proteins					494:501	the coronavirus M proteins	476:501	the coronavirus M proteins	476:501	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	9	2	theme	SARS-CoV	1538:1545	arg1	protein					1550:1556	The SARS-CoV 3a protein	1534:1556	The SARS-CoV 3a protein	1534:1556	The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides.
16474139	11	3	theme	serine	1836:1841	arg1	substitution					1820:1831	substitution	1820:1831	substitution of serine and threonine residues in the ectodomain of the 3a protein	1820:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.
16474139	7	4	theme	different	1300:1308	arg1	ways					1310:1313	different ways	1300:1313	different ways	1300:1313	Pulse-chase analysis showed that both proteins were modified, although in different ways.
16474139	11	5	theme	3a	1891:1892	arg1	protein					1894:1900	the 3a protein	1887:1900	the 3a protein	1887:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.
16474139	1	6	theme	reading	177:183	arg1	frame					185:189	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame	111:189	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	10	7	dep	in	1719:1720	arg1	situ					1722:1725	situ	1722:1725	situ	1722:1725	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.
16474139	3	8	theme	membrane	524:531	arg1	spanning					533:540	triple membrane spanning	517:540	triple membrane spanning	517:540	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	3	8	theme	membrane	524:531	arg1	they					508:511	they	508:511	they	508:511	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	1	9	theme	3a	191:192	arg1	protein					245:251	a structural protein	232:251	a structural protein	232:251	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	1	9	theme	3a	191:192	arg1	protein					194:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	3	10	theme	viral	655:659	arg1	proteins					672:679	viral structural proteins	655:679	viral structural proteins	655:679	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	5	11	theme	conserved	927:935	arg1	glycosylation					937:949	The conserved glycosylation	923:949	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them	923:1025	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	4	12	theme	crucial	792:798	arg1	role					800:803	a crucial role	790:803	a crucial role	790:803	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.
16474139	12	13	theme	O-glycosylated	1985:1998	arg1	protein					1971:1977	the SARS-CoV 3a protein	1955:1977	the SARS-CoV 3a protein	1955:1977	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	12	13	theme	O-glycosylated	1985:1998	arg1	glycoprotein					2000:2011	an O-glycosylated glycoprotein	1982:2011	an O-glycosylated glycoprotein	1982:2011	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	3	14	theme	same	551:554	arg1	topology					556:563	the same topology	547:563	the same topology	547:563	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	9	15	theme	sialic	1596:1601	arg1	acids					1603:1607	sialic acids	1596:1607	sialic acids	1596:1607	The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides.
16474139	6	16	theme	T7	1172:1173	arg1	system					1186:1191	the vaccinia virus T7 expression system	1153:1191	the vaccinia virus T7 expression system	1153:1191	The proteins were expressed separately using the vaccinia virus T7 expression system, followed by metabolic labeling.
16474139	10	17	theme	3a	1678:1679	arg1	protein					1681:1687	the 3a protein	1674:1687	the 3a protein	1674:1687	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.
16474139	1	18	theme	acute	122:126	arg1	SARS-CoV					162:169	SARS-CoV	162:169	SARS-CoV	162:169	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	1	18	theme	acute	122:126	arg1	coronavirus					149:159	The severe acute respiratory syndrome coronavirus	111:159	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	4	19	theme	M	774:774	arg1	protein					776:782	The M protein	770:782	The M protein	770:782	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.
16474139	12	20	theme	3a	1968:1969	arg1	protein					1971:1977	the SARS-CoV 3a protein	1955:1977	the SARS-CoV 3a protein	1955:1977	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	12	20	theme	3a	1968:1969	arg1	glycoprotein					2000:2011	an O-glycosylated glycoprotein	1982:2011	an O-glycosylated glycoprotein	1982:2011	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	6	21	theme	vaccinia	1157:1164	arg1	system					1186:1191	the vaccinia virus T7 expression system	1153:1191	the vaccinia virus T7 expression system	1153:1191	The proteins were expressed separately using the vaccinia virus T7 expression system, followed by metabolic labeling.
16474139	1	22	theme	syndrome	140:147	arg1	SARS-CoV					162:169	SARS-CoV	162:169	SARS-CoV	162:169	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	1	22	theme	syndrome	140:147	arg1	coronavirus					149:159	The severe acute respiratory syndrome coronavirus	111:159	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	3	23	theme	S	730:730	arg1	proteins					732:739	the E and S proteins	720:739	proteins	732:739	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	0	24	theme	triple-spanning	67:81	arg1	proteins					92:99	coronavirus triple-spanning membrane proteins 3a and M	55:108	coronavirus triple-spanning membrane proteins 3a and M	55:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	0	24	theme	triple-spanning	67:81	arg1	M					108:108	M	108:108	M	108:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	0	24	theme	triple-spanning	67:81	arg1	3a					101:102	3a	101:102	3a	101:102	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	3	25	theme	E	724:724	arg1	proteins					732:739	the E and S proteins	720:739	proteins	732:739	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	5	26	theme	coronavirus	958:968	arg1	proteins					972:979	the coronavirus M proteins	954:979	the coronavirus M proteins	954:979	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	3	27	contain	have	447:450	arg1	It					429:430	It	429:430	It	429:430	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	3	27	contain	have	447:450	arg2	similarities					460:471	several similarities	452:471	several similarities to the coronavirus M proteins	452:501	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	2	28	theme	known	367:371	arg1	proteins					402:409	the known structural or group-specific proteins	363:409	the known structural or group-specific proteins of coronaviruses	363:426	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.
16474139	6	29	theme	metabolic	1206:1214	arg1	labeling					1216:1223	metabolic labeling	1206:1223	metabolic labeling	1206:1223	The proteins were expressed separately using the vaccinia virus T7 expression system, followed by metabolic labeling.
16474139	5	30	theme	proteins	972:979	arg1	glycosylation					937:949	The conserved glycosylation	923:949	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them	923:1025	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	12	31	gly	glycosylated	2100:2111	arg1	protein					2080:2086	the SARS-CoV M protein	2065:2086	the SARS-CoV M protein	2065:2086	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	12	32	theme	M	2078:2078	arg1	protein					2080:2086	the SARS-CoV M protein	2065:2086	the SARS-CoV M protein	2065:2086	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	3	33	dep	Golgi	632:636	arg1	predominantly					618:630	predominantly	618:630	predominantly	618:630	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	0	34	dep	proteins	92:99	arg1	proteins					92:99	coronavirus triple-spanning membrane proteins 3a and M	55:108	coronavirus triple-spanning membrane proteins 3a and M	55:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	0	34	dep	proteins	92:99	arg1	M					108:108	M	108:108	M	108:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	0	34	dep	proteins	92:99	arg1	Glycosylation					0:12	Glycosylation	0:12	Glycosylation of the severe acute respiratory syndrome	0:53	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	0	34	dep	proteins	92:99	arg1	3a					101:102	3a	101:102	3a	101:102	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	2	35	theme	coronaviruses	414:426	arg1	proteins					402:409	the known structural or group-specific proteins	363:409	the known structural or group-specific proteins of coronaviruses	363:426	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.
16474139	8	36	theme	3a	1418:1419	arg1	protein					1421:1427	the 3a protein	1414:1427	the 3a protein	1414:1427	While the M protein acquired cotranslationally oligosaccharides that could be removed by PNGaseF, the 3a protein acquired its modifications posttranslationally, and they were not sensitive to the N-glycosidase enzyme.
16474139	11	37	dep	serine	1836:1841	arg1	residues					1857:1864	residues	1857:1864	residues	1857:1864	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.
16474139	12	38	dep	glycosylated	2100:2111	arg1	N					2098:2098	N	2098:2098	N	2098:2098	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	3	39	theme	similar	581:587	arg1	localizations					603:615	similar intracellular localizations	581:615	similar intracellular localizations	581:615	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	3	40	dep	spanning	533:540	arg1	i					505:505	i	505:505	i	505:505	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	3	40	dep	spanning	533:540	arg1	both					646:649	both	646:649	both	646:649	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	5	41	theme	3a	1008:1009	arg1	protein					1011:1017	the 3a protein	1004:1017	the 3a protein	1004:1017	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	0	42	theme	acute	28:32	arg1	syndrome					46:53	the severe acute respiratory syndrome	17:53	the severe acute respiratory syndrome	17:53	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	9	43	contain	contain	1588:1594	arg2	acids					1603:1607	sialic acids	1596:1607	sialic acids	1596:1607	The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides.
16474139	9	43	contain	contain	1588:1594	arg1	protein					1550:1556	The SARS-CoV 3a protein	1534:1556	The SARS-CoV 3a protein	1534:1556	The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides.
16474139	0	44	theme	syndrome	46:53	arg1	Glycosylation					0:12	Glycosylation	0:12	Glycosylation of the severe acute respiratory syndrome	0:53	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	5	45	gly	glycosylation	937:949	arg1	proteins					972:979	the coronavirus M proteins	954:979	the coronavirus M proteins	954:979	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	5	45	gly	glycosylation	937:949	arg1	resemblance					989:999	the resemblance	985:999	the resemblance of the 3a protein to them	985:1025	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	10	46	theme	mutants	1783:1789	arg1	assay					1743:1747	an in situ O-glycosylation assay	1716:1747	an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants	1716:1789	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.
16474139	2	47	theme	genes	316:320	arg1	genes					316:320	the so-called group-specific genes	287:320	the so-called group-specific genes	287:320	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.
16474139	2	47	theme	genes	316:320	arg1	one					280:282	one	280:282	one	280:282	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.
16474139	10	48	theme	endoplasmic	1752:1762	arg1	mutants					1783:1789	endoplasmic reticulum-retained mutants	1752:1789	endoplasmic reticulum-retained mutants	1752:1789	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.
16474139	12	49	theme	M	2043:2043	arg1	proteins					2045:2052	the group 2 coronavirus M proteins	2019:2052	the group 2 coronavirus M proteins	2019:2052	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	3	50	theme	M	492:492	arg1	proteins					494:501	the coronavirus M proteins	476:501	the coronavirus M proteins	476:501	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	3	51	dep	have	576:579	arg1	Golgi					632:636	Golgi	632:636	Golgi	632:636	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	3	51	dep	have	576:579	arg1	iii					641:643	iii	641:643	iii	641:643	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	3	51	dep	have	576:579	arg1	ii					567:568	ii	567:568	ii	567:568	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	2	52	theme	sequence	333:340	arg1	homology					342:349	no sequence homology	330:349	no sequence homology with any of the known structural or group-specific proteins of coronaviruses	330:426	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.
16474139	1	53	theme	structural	234:243	arg1	protein					245:251	a structural protein	232:251	a structural protein	232:251	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	1	53	theme	structural	234:243	arg1	protein					194:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	11	54	theme	protein	1894:1900	arg1	protein					1894:1900	the 3a protein	1887:1900	the 3a protein	1887:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.
16474139	11	54	theme	protein	1894:1900	arg1	ectodomain					1873:1882	the ectodomain	1869:1882	the ectodomain of the 3a protein	1869:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.
16474139	2	55	theme	so-called	291:299	arg1	genes					316:320	the so-called group-specific genes	287:320	the so-called group-specific genes	287:320	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.
16474139	9	56	theme	3a	1547:1548	arg1	protein					1550:1556	The SARS-CoV 3a protein	1534:1556	The SARS-CoV 3a protein	1534:1556	The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides.
16474139	8	57	theme	N-glycosidase	1512:1524	arg1	enzyme					1526:1531	the N-glycosidase enzyme	1508:1531	the N-glycosidase enzyme	1508:1531	While the M protein acquired cotranslationally oligosaccharides that could be removed by PNGaseF, the 3a protein acquired its modifications posttranslationally, and they were not sensitive to the N-glycosidase enzyme.
16474139	1	58	theme	open	172:175	arg1	frame					185:189	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame	111:189	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	3	59	theme	triple	517:522	arg1	spanning					533:540	triple membrane spanning	517:540	triple membrane spanning	517:540	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	3	59	theme	triple	517:522	arg1	they					508:511	they	508:511	they	508:511	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	1	60	theme	frame	185:189	arg1	protein					245:251	a structural protein	232:251	a structural protein	232:251	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	1	60	theme	frame	185:189	arg1	protein					194:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	4	61	link	O-linked	896:903	arg1	oligosaccharides					905:920	O-linked oligosaccharides	896:920	O-linked oligosaccharides	896:920	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.
16474139	3	62	theme	structural	661:670	arg1	proteins					672:679	viral structural proteins	655:679	viral structural proteins	655:679	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	3	63	dep	appear	696:701	arg1	iv					687:688	iv	687:688	iv	687:688	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	11	64	theme	threonine	1847:1855	arg1	substitution					1820:1831	substitution	1820:1831	substitution of serine and threonine residues in the ectodomain of the 3a protein	1820:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.
16474139	11	65	theme	sugars	1941:1946	arg1	addition					1916:1923	the addition	1912:1923	the addition of the O-linked sugars	1912:1946	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.
16474139	8	66	theme	M	1326:1326	arg1	protein					1328:1334	the M protein	1322:1334	the M protein	1322:1334	While the M protein acquired cotranslationally oligosaccharides that could be removed by PNGaseF, the 3a protein acquired its modifications posttranslationally, and they were not sensitive to the N-glycosidase enzyme.
16474139	3	67	contain	have	576:579	arg2	localizations					603:615	similar intracellular localizations	581:615	similar intracellular localizations	581:615	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	3	67	contain	have	576:579	arg1	they					571:574	they	571:574	they	571:574	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	6	68	theme	expression	1175:1184	arg1	system					1186:1191	the vaccinia virus T7 expression system	1153:1191	the vaccinia virus T7 expression system	1153:1191	The proteins were expressed separately using the vaccinia virus T7 expression system, followed by metabolic labeling.
16474139	10	69	theme	protein	1681:1687	arg1	O-glycosylation					1655:1669	O-glycosylation	1655:1669	O-glycosylation of the 3a protein	1655:1687	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.
16474139	1	70	theme	severe	115:120	arg1	SARS-CoV					162:169	SARS-CoV	162:169	SARS-CoV	162:169	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	1	70	theme	severe	115:120	arg1	coronavirus					149:159	The severe acute respiratory syndrome coronavirus	111:159	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	11	71	from	substitution	1820:1831	arg1	protein					1894:1900	the 3a protein	1887:1900	the 3a protein	1887:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.
16474139	11	71	from	substitution	1820:1831	arg1	ectodomain					1873:1882	the ectodomain	1869:1882	the ectodomain of the 3a protein	1869:1900	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.
16474139	6	72	theme	virus	1166:1170	arg1	system					1186:1191	the vaccinia virus T7 expression system	1153:1191	the vaccinia virus T7 expression system	1153:1191	The proteins were expressed separately using the vaccinia virus T7 expression system, followed by metabolic labeling.
16474139	10	73	theme	in	1719:1720	arg1	assay					1743:1747	an in situ O-glycosylation assay	1716:1747	an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants	1716:1789	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.
16474139	1	74	theme	respiratory	128:138	arg1	SARS-CoV					162:169	SARS-CoV	162:169	SARS-CoV	162:169	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	1	74	theme	respiratory	128:138	arg1	coronavirus					149:159	The severe acute respiratory syndrome coronavirus	111:159	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	7	75	theme	Pulse-chase	1226:1236	arg1	analysis					1238:1245	Pulse-chase analysis	1226:1245	Pulse-chase analysis	1226:1245	Pulse-chase analysis showed that both proteins were modified, although in different ways.
16474139	12	76	theme	SARS-CoV	1959:1966	arg1	protein					1971:1977	the SARS-CoV 3a protein	1955:1977	the SARS-CoV 3a protein	1955:1977	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	12	76	theme	SARS-CoV	1959:1966	arg1	glycoprotein					2000:2011	an O-glycosylated glycoprotein	1982:2011	an O-glycosylated glycoprotein	1982:2011	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	0	77	theme	membrane	83:90	arg1	proteins					92:99	coronavirus triple-spanning membrane proteins 3a and M	55:108	coronavirus triple-spanning membrane proteins 3a and M	55:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	0	77	theme	membrane	83:90	arg1	M					108:108	M	108:108	M	108:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	0	77	theme	membrane	83:90	arg1	3a					101:102	3a	101:102	3a	101:102	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	12	78	gly	O-glycosylated	1985:1998	arg1	protein					1971:1977	the SARS-CoV 3a protein	1955:1977	the SARS-CoV 3a protein	1955:1977	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	12	78	gly	O-glycosylated	1985:1998	arg1	glycoprotein					2000:2011	an O-glycosylated glycoprotein	1982:2011	an O-glycosylated glycoprotein	1982:2011	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	1	79	theme	coronavirus	149:159	arg1	frame					185:189	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame	111:189	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein	111:200	The severe acute respiratory syndrome coronavirus (SARS-CoV) open reading frame 3a protein has recently been shown to be a structural protein.
16474139	5	80	theme	resemblance	989:999	arg1	glycosylation					937:949	The conserved glycosylation	923:949	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them	923:1025	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	9	81	theme	oligosaccharides	1637:1652	arg1	presence					1625:1632	the presence	1621:1632	the presence of oligosaccharides	1621:1652	The SARS-CoV 3a protein, however, was demonstrated to contain sialic acids, indicating the presence of oligosaccharides.
16474139	10	82	theme	reticulum-retained	1764:1781	arg1	mutants					1783:1789	endoplasmic reticulum-retained mutants	1752:1789	endoplasmic reticulum-retained mutants	1752:1789	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.
16474139	12	83	gly	glycoprotein	2000:2011	arg1	protein					1971:1977	the SARS-CoV 3a protein	1955:1977	the SARS-CoV 3a protein	1955:1977	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	12	83	gly	glycoprotein	2000:2011	arg1	glycoprotein					2000:2011	an O-glycosylated glycoprotein	1982:2011	an O-glycosylated glycoprotein	1982:2011	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	2	84	contain	has	326:328	arg1	protein					258:264	The protein	254:264	The protein	254:264	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.
16474139	2	84	contain	has	326:328	arg2	homology					342:349	no sequence homology	330:349	no sequence homology with any of the known structural or group-specific proteins of coronaviruses	330:426	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.
16474139	3	85	theme	several	452:458	arg1	similarities					460:471	several similarities	452:471	several similarities to the coronavirus M proteins	452:501	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	5	86	theme	M	970:970	arg1	proteins					972:979	the coronavirus M proteins	954:979	the coronavirus M proteins	954:979	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	4	87	gly	glycosylated	836:847	arg1	coronaviruses					856:868	all coronaviruses	852:868	all coronaviruses	852:868	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.
16474139	4	87	gly	glycosylated	836:847	arg1	protein					776:782	The M protein	770:782	The M protein	770:782	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.
16474139	4	87	gly	glycosylated	836:847	arg2	protein					776:782	The M protein	770:782	The M protein	770:782	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.
16474139	11	88	link	O-linked	1932:1939	arg1	sugars					1941:1946	the O-linked sugars	1928:1946	the O-linked sugars	1928:1946	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.
16474139	4	89	theme	coronavirus	808:818	arg1	assembly					820:827	coronavirus assembly	808:827	coronavirus assembly	808:827	The M protein plays a crucial role in coronavirus assembly and is glycosylated in all coronaviruses, either by N-linked or by O-linked oligosaccharides.
16474139	5	90	gly	glycosylation	1053:1065	arg1	proteins					1098:1105	these two SARS-CoV membrane proteins	1070:1105	these two SARS-CoV membrane proteins	1070:1105	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	11	91	theme	O-linked	1932:1939	arg1	sugars					1941:1946	the O-linked sugars	1928:1946	the O-linked sugars	1928:1946	In addition, we showed that substitution of serine and threonine residues in the ectodomain of the 3a protein abolished the addition of the O-linked sugars.
16474139	12	92	theme	SARS-CoV	2069:2076	arg1	protein					2080:2086	the SARS-CoV M protein	2065:2086	the SARS-CoV M protein	2065:2086	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	0	93	theme	severe	21:26	arg1	syndrome					46:53	the severe acute respiratory syndrome	17:53	the severe acute respiratory syndrome	17:53	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	2	94	theme	group-specific	387:400	arg1	proteins					402:409	the known structural or group-specific proteins	363:409	the known structural or group-specific proteins of coronaviruses	363:426	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.
16474139	0	95	theme	respiratory	34:44	arg1	syndrome					46:53	the severe acute respiratory syndrome	17:53	the severe acute respiratory syndrome	17:53	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	10	96	gly	O-glycosylation	1655:1669	arg1	protein					1681:1687	the 3a protein	1674:1687	the 3a protein	1674:1687	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.
16474139	0	97	gly	Glycosylation	0:12	arg1	syndrome					46:53	the severe acute respiratory syndrome	17:53	the severe acute respiratory syndrome	17:53	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	10	98	theme	O-glycosylation	1727:1741	arg1	assay					1743:1747	an in situ O-glycosylation assay	1716:1747	an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants	1716:1789	O-glycosylation of the 3a protein was indeed confirmed using an in situ O-glycosylation assay of endoplasmic reticulum-retained mutants.
16474139	5	99	theme	membrane	1089:1096	arg1	proteins					1098:1105	these two SARS-CoV membrane proteins	1070:1105	these two SARS-CoV membrane proteins	1070:1105	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	2	100	theme	structural	373:382	arg1	proteins					402:409	the known structural or group-specific proteins	363:409	the known structural or group-specific proteins of coronaviruses	363:426	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.
16474139	0	101	theme	coronavirus	55:65	arg1	proteins					92:99	coronavirus triple-spanning membrane proteins 3a and M	55:108	coronavirus triple-spanning membrane proteins 3a and M	55:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	0	101	theme	coronavirus	55:65	arg1	M					108:108	M	108:108	M	108:108	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	0	101	theme	coronavirus	55:65	arg1	3a					101:102	3a	101:102	3a	101:102	Glycosylation of the severe acute respiratory syndrome coronavirus triple-spanning membrane proteins 3a and M.
16474139	5	102	theme	protein	1011:1017	arg1	proteins					972:979	the coronavirus M proteins	954:979	the coronavirus M proteins	954:979	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	5	102	theme	protein	1011:1017	arg1	resemblance					989:999	the resemblance	985:999	the resemblance of the 3a protein to them	985:1025	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	3	103	theme	intracellular	589:601	arg1	localizations					603:615	similar intracellular localizations	581:615	similar intracellular localizations	581:615	It does, however, have several similarities to the coronavirus M proteins; (i) they are triple membrane spanning with the same topology, (ii) they have similar intracellular localizations (predominantly Golgi), (iii) both are viral structural proteins, and (iv) they appear to interact with the E and S proteins, as well as with each other.
16474139	12	104	theme	group	2023:2027	arg1	proteins					2045:2052	the group 2 coronavirus M proteins	2019:2052	the group 2 coronavirus M proteins	2019:2052	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	2	105	theme	group-specific	301:314	arg1	genes					316:320	the so-called group-specific genes	287:320	the so-called group-specific genes	287:320	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.
16474139	5	106	theme	SARS-CoV	1080:1087	arg1	proteins					1098:1105	these two SARS-CoV membrane proteins	1070:1105	these two SARS-CoV membrane proteins	1070:1105	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.
16474139	2	107	with	homology	342:349	arg1	any					356:358	any	356:358	any	356:358	The protein is encoded by one of the so-called group-specific genes and has no sequence homology with any of the known structural or group-specific proteins of coronaviruses.
16474139	12	108	theme	coronavirus	2031:2041	arg1	proteins					2045:2052	the group 2 coronavirus M proteins	2019:2052	the group 2 coronavirus M proteins	2019:2052	Thus, the SARS-CoV 3a protein is an O-glycosylated glycoprotein, like the group 2 coronavirus M proteins but unlike the SARS-CoV M protein, which is N glycosylated.
16474139	5	109	theme	proteins	1098:1105	arg1	glycosylation					1053:1065	the glycosylation	1049:1065	the glycosylation of these two SARS-CoV membrane proteins	1049:1105	The conserved glycosylation of the coronavirus M proteins and the resemblance of the 3a protein to them led us to investigate the glycosylation of these two SARS-CoV membrane proteins.