PMID- 10361113
OWN - NLM
STAT- MEDLINE
DCOM- 19990701
LR  - 20210216
IS  - 0006-4971 (Print)
IS  - 0006-4971 (Linking)
VI  - 93
IP  - 12
DP  - 1999 Jun 15
TI  - Dominant negative mutants implicate STAT5 in myeloid cell proliferation and
      neutrophil differentiation.
PG  - 4154-66
AB  - STAT5 is a member of the signal transducers and activation of transcription
      (STAT) family of latent transcription factors activated in a variety of cytokine 
      signaling pathways. We introduced alanine substitution mutations in highly
      conserved regions of murine STAT5A and studied the mutants for dimerization, DNA 
      binding, transactivation, and dominant negative effects on erythropoietin-induced
      STAT5-dependent transcriptional activation. The mutations included two near the
      amino-terminus (W255KR-->AAA and R290QQ-->AAA), two in the DNA-binding domain
      (E437E-->AA and V466VV-->AAA), and a carboxy-terminal truncation of STAT5A
      (STAT5A/triangle up53C) analogous to a naturally occurring isoform of rat STAT5B.
      All of the STAT mutant proteins were tyrosine phosphorylated by JAK2 and
      heterodimerized with STAT5B except for the WKR mutant, suggesting an important
      role for this region in STAT5 for stabilizing dimerization. The WKR, EE, and VVV 
      mutants had no detectable DNA-binding activity, and the WKR and VVV mutants, but 
      not EE, were defective in transcriptional induction. The VVV mutant had a
      moderate dominant negative effect on erythropoietin-induced STAT5 transcriptional
      activation, which was likely due to the formation of heterodimers that are
      defective in DNA binding. Interestingly, the WKR mutant had a potent dominant
      negative effect, comparable to the transactivation domain deletion mutant,
      triangle up53C. Stable expression of either the WKR or triangle up53C STAT5
      mutants in the murine myeloid cytokine-dependent cell line 32D inhibited both
      interleukin-3-dependent proliferation and granulocyte colony-stimulating factor
      (G-CSF)-dependent differentiation, without induction of apoptosis. Expression of 
      these mutants in primary murine bone marrow inhibited G-CSF-dependent granulocyte
      colony formation in vitro. These results demonstrate that mutations in distinct
      regions of STAT5 exert dominant negative effects on cytokine signaling, likely
      through different mechanisms, and suggest a role for STAT5 in proliferation and
      differentiation of myeloid cells.
FAU - Ilaria, R L Jr
AU  - Ilaria RL Jr
AD  - The Simmons Cancer Center, University of Texas Southwestern Medical School,
      Dallas, TX, USA.
FAU - Hawley, R G
AU  - Hawley RG
FAU - Van Etten, R A
AU  - Van Etten RA
LA  - eng
GR  - CA57593/CA/NCI NIH HHS/United States
GR  - HL03310/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Blood
JT  - Blood
JID - 7603509
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Milk Proteins)
RN  - 0 (Proto-Oncogene Proteins)
RN  - 0 (STAT5 Transcription Factor)
RN  - 0 (Stat5a protein, mouse)
RN  - 0 (Stat5a protein, rat)
RN  - 0 (Stat5b protein, mouse)
RN  - 0 (Stat5b protein, rat)
RN  - 0 (Trans-Activators)
RN  - 11096-26-7 (Erythropoietin)
RN  - 143011-72-7 (Granulocyte Colony-Stimulating Factor)
RN  - 21820-51-9 (Phosphotyrosine)
RN  - 9007-49-2 (DNA)
RN  - EC 2.7.10.1 (Protein-Tyrosine Kinases)
RN  - EC 2.7.10.2 (Jak2 protein, mouse)
RN  - EC 2.7.10.2 (Jak2 protein, rat)
RN  - EC 2.7.10.2 (Janus Kinase 2)
SB  - AIM
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - *Cell Differentiation
MH  - *Cell Division
MH  - Cell Line
MH  - DNA/metabolism
MH  - DNA-Binding Proteins/chemistry/genetics/*physiology
MH  - Dimerization
MH  - Erythropoietin/pharmacology
MH  - Granulocyte Colony-Stimulating Factor/pharmacology
MH  - Granulocytes/*cytology
MH  - Janus Kinase 2
MH  - Mice
MH  - *Milk Proteins
MH  - Molecular Sequence Data
MH  - *Mutagenesis
MH  - Neutrophils/*cytology
MH  - Phosphotyrosine/metabolism
MH  - Point Mutation
MH  - Protein-Tyrosine Kinases/metabolism
MH  - *Proto-Oncogene Proteins
MH  - Rats
MH  - STAT5 Transcription Factor
MH  - Trans-Activators/chemistry/genetics/*physiology
MH  - Transcriptional Activation
EDAT- 1999/06/11 10:00
MHDA- 2001/03/28 10:01
CRDT- 1999/06/11 10:00
PHST- 1999/06/11 10:00 [pubmed]
PHST- 2001/03/28 10:01 [medline]
PHST- 1999/06/11 10:00 [entrez]
AID - S0006-4971(20)59445-7 [pii]
PST - ppublish
SO  - Blood. 1999 Jun 15;93(12):4154-66.