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PMID: 17018296 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Cdc7-Dbf4 phosphorylates MCM proteins via a docking site-mediated mechanism to promote S phase progression.

Molecular cell ·Vol. 24 ·No. 1 ·2006-10-06 ·Pages 101-13

Sheu YJ, Stillman B

Abstract

Origins of DNA replication are licensed in G1 by recruiting the minichromosome maintenance (MCM) proteins to form a prereplicative complex (pre-RC). Prior to initiation of DNA synthesis from each origin, a preinitiation complex (pre-IC) containing Cdc45 and other proteins is formed. We report that Cdc7-Dbf4 protein kinase (DDK) promotes assembly of a stable Cdc45-MCM complex exclusively on chromatin in S phase. In this complex, Mcm4 is hyperphosphorylated. Studies in vitro using purified DDK and Mcm4 demonstrate that hyperphosphorylation occurs at the Mcm4 N terminus. However, the DDK substrate specificity is conferred by an adjacent DDK-docking domain (DDD), sufficient for facilitating efficient phosphorylation of artificial phosphoacceptors in cis. Genetic evidence suggests that phosphorylation of Mcm4 by DDK is important for timely S phase progression and for cell viability upon overproduction of Cdc45. We suggest that DDK docks on and phosphorylates MCM proteins at licensed origins to promote proper assembly of pre-IC.

MeSH Terms
Amino Acid Motifs Apoenzymes/metabolism Binding Sites Cell Cycle Proteins/chemistry,metabolism,physiology Chromatin/metabolism Conserved Sequence DNA Replication DNA-Binding Proteins/chemistry,metabolism,physiology Minichromosome Maintenance Complex Component 4 Models, Biological Molecular Sequence Data Nuclear Proteins/chemistry,metabolism,physiology Phosphorylation Protein Serine-Threonine Kinases/chemistry,metabolism,physiology Protein Structure, Tertiary S Phase/physiology Saccharomyces cerevisiae/cytology,genetics,metabolism Saccharomyces cerevisiae Proteins/chemistry,metabolism,physiology Sequence Alignment
Chemicals
Apoenzymes CDC45 protein, S cerevisiae Cell Cycle Proteins Chromatin DNA-Binding Proteins Dbf4 protein, S cerevisiae Nuclear Proteins Saccharomyces cerevisiae Proteins CDC7 protein, S cerevisiae Protein Serine-Threonine Kinases MCM4 protein, S cerevisiae Minichromosome Maintenance Complex Component 4
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Sheu Yi-Jun
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
Stillman Bruce
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Article Info
Journal
Molecular cell
Abbr.
Mol Cell
ISSN
1097-2765
DOI
10.1016/j.molcel.2006.07.033
Published
2006-10-06
Pages
101-13
Language
English
Region
United States
NLM ID
9802571
PMCID
PMC2923825
Subset
IM
Grants
NIGMS NIH HHS · R01 GM045436 · United States
NIGMS NIH HHS · R01 GM045436-18 · United States
NIGMS NIH HHS · GM 45436 · United States
External Links
PubMed source DOI: 10.1016/j.molcel.2006.07.033 PMC Full Text View journal details
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