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PMID: 10465792 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A role for the Cdc7 kinase regulatory subunit Dbf4p in the formation of initiation-competent origins of replication.

Genes & development ·Vol. 13 ·No. 16 ·1999-08-15 ·Pages 2159-76

Pasero P, Duncker BP, Schwob E, Gasser SM

Abstract

Using a reconstituted DNA replication assay from yeast, we demonstrate that two kinase complexes are essential for the promotion of replication in vitro. An active Clb/Cdc28 kinase complex, or its vertebrate equivalent, is required in trans to stimulate initiation in G(1)-phase nuclei, whereas the Dbf4/Cdc7 kinase complex must be provided by the template nuclei themselves. The regulatory subunit of Cdc7p, Dbf4p, accumulates during late G(1) phase, becomes chromatin associated prior to Clb/Cdc28 activation, and assumes a punctate pattern of localization that is similar to, and dependent on, the origin recognition complex (ORC). The association of Dbf4p with a detergent-insoluble chromatin fraction in G(1)-phase nuclei requires ORC but not Cdc6p or Clb/Cdc28 kinase activity, and correlates with competence for initiation. We propose a model in which Dbf4p targets Cdc7p to the prereplication complex prior to the G(1)/S transition, by a pathway parallel to, but independent of, the Cdc6p-dependent recruitment of MCMs.

MeSH Terms
CDC28 Protein Kinase, S cerevisiae/metabolism Cell Cycle Cell Cycle Proteins/metabolism Cell Fractionation Cell Nucleus/metabolism Chromatin/metabolism DNA, Fungal/biosynthesis DNA-Binding Proteins/metabolism Fungal Proteins/physiology G1 Phase Origin Recognition Complex Protein Serine-Threonine Kinases/metabolism Replication Origin Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins
Chemicals
CDC6 protein, S cerevisiae Cell Cycle Proteins Chromatin DNA, Fungal DNA-Binding Proteins Dbf4 protein, S cerevisiae Fungal Proteins Origin Recognition Complex Saccharomyces cerevisiae Proteins CDC7 protein, S cerevisiae Protein Serine-Threonine Kinases CDC28 Protein Kinase, S cerevisiae
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Pasero P
Swiss Institute for Experimental Cancer Research (ISREC), CH-1066 Epalinges/Lausanne, Switzerland.
Duncker B P
Schwob E
Gasser S M
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.13.16.2159
Published
1999-08-15
Pages
2159-76
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC316966
Subset
IM
External Links
PubMed source DOI: 10.1101/gad.13.16.2159 PMC Full Text View journal details
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