PMID- 10523313
OWN - NLM
STAT- MEDLINE
DCOM- 19991206
LR  - 20161019
IS  - 0261-4189 (Print)
IS  - 0261-4189 (Linking)
VI  - 18
IP  - 20
DP  - 1999 Oct 15
TI  - Mammalian Cdc7-Dbf4 protein kinase complex is essential for initiation of DNA
      replication.
PG  - 5703-13
AB  - The Cdc7-Dbf4 kinase is essential for regulating initiation of DNA replication in
      Saccharomyces cerevisiae. Previously, we identified a human Cdc7 homolog, HsCdc7.
      In this study, we report the identification of a human Dbf4 homolog, HsDbf4. We
      show that HsDbf4 binds to HsCdc7 and activates HsCdc7 kinase activity when HsDbf4
      and HsCdc7 are coexpressed in insect and mammalian cells. HsDbf4 protein levels
      are regulated during the cell cycle with a pattern that matches that of HsCdc7
      protein kinase activity. They are low in G(1), increase during G(1)-S, and remain
      high during S and G(2)-M. Purified baculovirus-expressed HsCdc7-HsDbf4
      selectively phosphorylates the MCM2 subunit of the minichromosome maintenance
      (MCM) protein complex isolated by immunoprecipitation with MCM7 antibodies in
      vitro. Two-dimensional tryptic phosphopeptide-mapping analysis of in vivo
      (32)P-labeled MCM2 from HeLa cells reveals that several major tryptic
      phosphopeptides of MCM2 comigrate with those of MCM2 phosphorylated by
      HsCdc7-HsDbf4 in vitro, suggesting that MCM2 is a physiological HsCdc7-HsDbf4
      substrate. Immunoneutralization of HsCdc7-HsDbf4 activity by microinjection of
      anti-HsCdc7 antibodies into HeLa cells blocks initiation of DNA replication.
      These results indicate that the HsCdc7-HsDbf4 kinase is directly involved in
      regulating the initiation of DNA replication by targeting MCM2 protein in
      mammalian cells.
FAU - Jiang, W
AU  - Jiang W
AD  - The Salk Institute, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA.
      wjiang@salk.edu
FAU - McDonald, D
AU  - McDonald D
FAU - Hope, T J
AU  - Hope TJ
FAU - Hunter, T
AU  - Hunter T
LA  - eng
SI  - GENBANK/AF160249
GR  - CA14195/CA/NCI NIH HHS/United States
GR  - CA39780/CA/NCI NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - EMBO J
JT  - The EMBO journal
JID - 8208664
RN  - 0 (Cell Cycle Proteins)
RN  - 0 (DBF4 protein, human)
RN  - 0 (Multienzyme Complexes)
RN  - 0 (Nuclear Proteins)
RN  - 0 (Saccharomyces cerevisiae Proteins)
RN  - EC 2.7.1.- (CDC7 protein, S cerevisiae)
RN  - EC 2.7.1.- (CDC7 protein, human)
RN  - EC 2.7.1.- (Cdc7 protein, mouse)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
RN  - EC 3.6.4.12 (Minichromosome Maintenance Complex Component 2)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Cell Cycle
MH  - Cell Cycle Proteins/chemistry/genetics/*metabolism
MH  - Cell Line
MH  - Cloning, Molecular
MH  - DNA Replication/genetics/*physiology
MH  - Enzyme Activation
MH  - Humans
MH  - In Vitro Techniques
MH  - Minichromosome Maintenance Complex Component 2
MH  - Molecular Sequence Data
MH  - Multienzyme Complexes/chemistry/genetics/metabolism
MH  - Nuclear Proteins/metabolism
MH  - Phosphorylation
MH  - Protein-Serine-Threonine Kinases/chemistry/genetics/*metabolism
MH  - Saccharomyces cerevisiae/genetics/metabolism
MH  - *Saccharomyces cerevisiae Proteins
MH  - Substrate Specificity
PMC - PMC1171637
EDAT- 1999/10/16 00:00
MHDA- 1999/10/16 00:01
CRDT- 1999/10/16 00:00
PHST- 1999/10/16 00:00 [pubmed]
PHST- 1999/10/16 00:01 [medline]
PHST- 1999/10/16 00:00 [entrez]
AID - 10.1093/emboj/18.20.5703 [doi]
PST - ppublish
SO  - EMBO J. 1999 Oct 15;18(20):5703-13. doi: 10.1093/emboj/18.20.5703.