PMID- 10069453
OWN - NLM
STAT- MEDLINE
DCOM- 19990311
LR  - 20190513
IS  - 0143-3334 (Print)
IS  - 0143-3334 (Linking)
VI  - 20
IP  - 2
DP  - 1999 Feb
TI  - Defining the substrate specificity of cdk4 kinase-cyclin D1 complex.
PG  - 193-8
AB  - cdk4 kinase-cyclin D1 complex (cdk4/D1) does not phosphorylate all of the sites
      within retinoblastoma protein (Rb) equally. Comparison of five phosphorylation
      sites within the 15 kDa C domain of Rb indicates that Ser795 is the preferred
      site of phosphorylation by cdk4/D1. A series of experiments has been performed to
      determine the properties of this site that direct preferential phosphorylation.
      For cdk4/D1, the preferred amino acid at the third position C-terminal to the
      phosphorylated serine/threonine is arginine. Substitution of other amino acids,
      including a conservative change to lysine, has dramatic effects on the rates of
      phosphorylation. This information has been used to mutate less favorable sites in
      Rb, converting them to sites that are now preferentially phosphorylated by
      cdk4/D1. A conserved site at Ser842 in the related pocket protein p107 is also
      preferentially phosphorylated by cdk4/D1. Although Rb and p107 differ
      significantly in sequence, the Rb Ser795 site can replace the p107 Ser842 site
      without affecting the rate of phosphorylation. These results suggest that
      although a determinant of specificity resides in the sequences surrounding the
      phosphorylated site, the structural context of the site is also a critical
      parameter of specificity.
FAU - Grafstrom, R H
AU  - Grafstrom RH
AD  - Genetics and Cancer Group, Dupont Pharmaceutical Co., Wilmington, DE 19880-0336, 
      USA. robert.h.grafstrom@dupontpharma.com
FAU - Pan, W
AU  - Pan W
FAU - Hoess, R H
AU  - Hoess RH
LA  - eng
PT  - Journal Article
PL  - England
TA  - Carcinogenesis
JT  - Carcinogenesis
JID - 8008055
RN  - 0 (Nuclear Proteins)
RN  - 0 (Proto-Oncogene Proteins)
RN  - 0 (RBL1 protein, human)
RN  - 0 (Retinoblastoma Protein)
RN  - 0 (Retinoblastoma-Like Protein p107)
RN  - 136601-57-5 (Cyclin D1)
RN  - 2ZD004190S (Threonine)
RN  - 452VLY9402 (Serine)
RN  - 94ZLA3W45F (Arginine)
RN  - EC 2.7.11.22 (CDK4 protein, human)
RN  - EC 2.7.11.22 (Cyclin-Dependent Kinase 4)
RN  - EC 2.7.11.22 (Cyclin-Dependent Kinases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Arginine/metabolism
MH  - Binding Sites
MH  - Conserved Sequence
MH  - Cyclin D1/*metabolism
MH  - Cyclin-Dependent Kinase 4
MH  - Cyclin-Dependent Kinases/*metabolism
MH  - HeLa Cells
MH  - Humans
MH  - Nuclear Proteins/*metabolism
MH  - Phosphorylation
MH  - *Proto-Oncogene Proteins
MH  - Retinoblastoma Protein/*metabolism
MH  - Retinoblastoma-Like Protein p107
MH  - Serine/metabolism
MH  - Substrate Specificity
MH  - Threonine/metabolism
EDAT- 1999/03/09 00:00
MHDA- 1999/03/09 00:01
CRDT- 1999/03/09 00:00
PHST- 1999/03/09 00:00 [pubmed]
PHST- 1999/03/09 00:01 [medline]
PHST- 1999/03/09 00:00 [entrez]
AID - 10.1093/carcin/20.2.193 [doi]
PST - ppublish
SO  - Carcinogenesis. 1999 Feb;20(2):193-8. doi: 10.1093/carcin/20.2.193.