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Titolo:
Mammalian Cdc7-Dbf4 protein kinase complex is essential for initiation of DNA replication
Autore:
Jiang, W; McDonald, D; Hope, TJ; Hunter, T;
Indirizzi:
Salk Inst, La Jolla, CA 92037 USA Salk Inst La Jolla CA USA 92037Salk Inst, La Jolla, CA 92037 USA
Titolo Testata:
EMBO JOURNAL
fascicolo: 20, volume: 18, anno: 1999,
pagine: 5703 - 5713
SICI:
0261-4189(19991015)18:20<5703:MCPKCI>2.0.ZU;2-7
Fonte:
ISI
Lingua:
ENG
Soggetto:
MCM HETEROHEXAMERIC COMPLEXES; CELL-CYCLE; SACCHAROMYCES-CEREVISIAE; BUDDING YEAST; S-PHASE; HUMAN HOMOLOG; IN-VIVO; CDC7; CHROMATIN; PHOSPHORYLATION;
Keywords:
DNA replication; HsCdc7; HsDbf4; kinase; MCM;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
40
Recensione:
Indirizzi per estratti:
Indirizzo: Jiang, W NYU, Med Ctr, Dept Biochem, 550 1st Ave, New York, NY 10016 USA NYU 550 1st Ave New York NY USA 10016 Ave, New York, NY 10016 USA
Citazione:
W. Jiang et al., "Mammalian Cdc7-Dbf4 protein kinase complex is essential for initiation of DNA replication", EMBO J, 18(20), 1999, pp. 5703-5713

Abstract

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 byimmunoprecipitation with MCM7 antibodies in vitro, Two-dimensional trypticphosphopeptide-mapping analysis of in vivo P-32-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 HsCdc7HsDbf4 activity by microinjection of anti-HsCdc7 antibodies into HeLa cells blocks initiation of DNA replication. These results indicate that the HsCdc7HsDbf4 kinase is directly involved in regulating the initiation of DNAreplication by targeting MCM2 protein in mammalian cells.

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Documento generato il 22/01/20 alle ore 06:23:36