PMID- 10339564
OWN - NLM
STAT- MEDLINE
DCOM- 19990624
LR  - 20190503
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 96
IP  - 11
DP  - 1999 May 25
TI  - Multistep regulation of DNA replication by Cdk phosphorylation of HsCdc6.
PG  - 6193-8
AB  - We have characterized HsCdc6, a human protein homologous to the budding yeast
      Cdc6p that is essential for DNA replication. We show that, unlike Cdc6p, the
      levels of HsCdc6 protein remain constant throughout the cell cycle in human
      cells. However, phosphorylation of HsCdc6 is regulated during the cell cycle.
      HsCdc6 is an excellent substrate for Cdk2 in vitro and is phosphorylated in vivo 
      at three sites (Ser-54, Ser-74, and Ser-106) that are phosphorylated by Cdk2 in
      vitro, strongly suggesting that HsCdc6 is an in vivo Cdk substrate. HsCdc6 is
      nuclear in G1, but translocates to the cytoplasm at the start of S phase via
      Crm1-dependent export. An HsCdc6A1A2A3 mutant, which mimics unphosphorylated
      HsCdc6, is exclusively nuclear, and its expression inhibits initiation of DNA
      replication. An HsCdc6E1E2E3 mutant, which mimics phosphorylated HsCdc6, is
      exclusively cytoplasmic and is not associated with the chromatin/nuclear matrix
      fraction. Based on these results, we propose that phosphorylation of HsCdc6 by
      Cdks regulates DNA replication of at least two steps: first, by promoting
      initiation of DNA replication and, second, through nuclear exclusion preventing
      DNA rereplication.
FAU - Jiang, W
AU  - Jiang W
AD  - Molecular Biology and Virology Laboratory, The Salk Institute, 10010 North Torrey
      Pines Road, La Jolla, CA 92037, USA. wjiang@salk.edu
FAU - Wells, N J
AU  - Wells NJ
FAU - Hunter, T
AU  - Hunter T
LA  - eng
GR  - Wellcome Trust/United Kingdom
GR  - P30 CA014195/CA/NCI NIH HHS/United States
GR  - CA14195/CA/NCI NIH HHS/United States
GR  - CA39780/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (Antibodies)
RN  - 0 (CDC6 protein, S cerevisiae)
RN  - 0 (CDC6 protein, human)
RN  - 0 (Cell Cycle Proteins)
RN  - 0 (Chromatin)
RN  - 0 (Nuclear Proteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Saccharomyces cerevisiae Proteins)
RN  - EC 2.7.11.22 (Cyclin-Dependent Kinases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Antibodies
MH  - Cell Cycle/*physiology
MH  - Cell Cycle Proteins/chemistry/*metabolism
MH  - Cell Line
MH  - Cells, Cultured
MH  - Chromatin/physiology
MH  - Cyclin-Dependent Kinases/*metabolism
MH  - *DNA Replication
MH  - Fibroblasts/cytology/physiology
MH  - HeLa Cells
MH  - Humans
MH  - Infant, Newborn
MH  - Male
MH  - Molecular Sequence Data
MH  - Nuclear Matrix/physiology
MH  - Nuclear Proteins/chemistry/*metabolism
MH  - Peptide Fragments/chemistry/immunology
MH  - Phosphorylation
MH  - Recombinant Proteins/metabolism
MH  - Saccharomyces cerevisiae/metabolism
MH  - *Saccharomyces cerevisiae Proteins
MH  - Schizosaccharomyces/metabolism
MH  - Skin/cytology
PMC - PMC26858
EDAT- 1999/05/26 00:00
MHDA- 1999/05/26 00:01
CRDT- 1999/05/26 00:00
PHST- 1999/05/26 00:00 [pubmed]
PHST- 1999/05/26 00:01 [medline]
PHST- 1999/05/26 00:00 [entrez]
AID - 10.1073/pnas.96.11.6193 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 1999 May 25;96(11):6193-8. doi:
      10.1073/pnas.96.11.6193.