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PMID: 9670028 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Evidence for a Cdc6p-independent mitotic resetting event involving DNA polymerase alpha.

The EMBO journal ·Vol. 17 ·No. 14 ·1998-07-15 ·Pages 4139-46

Desdouets C, Santocanale C, Drury LS, Perkins G, Foiani M, Plevani P, Diffley JF

Abstract

Eukaryotic DNA replication is limited to once per cell cycle because cyclin-dependent kinases (cdks), which are required to fire origins, also prevent re-replication. Components of the replication apparatus, therefore, are 'reset' by cdk inactivation at the end of mitosis. In budding yeast, assembly of Cdc6p-dependent pre-replicative complexes (pre-RCs) at origins can only occur during G1 because it is blocked by cdk1 (Cdc28) together with B cyclins (Clbs). Here we describe a second, separate process which is also blocked by Cdc28/Clb kinase and, therefore, can only occur during G1; the recruitment of DNA polymerase alpha-primase (pol alpha) to chromatin. The recruitment of pol alpha to chromatin during G1 is independent of pre-RC formation since it can occur in the absence of Cdc6 protein. Paradoxically, overproduction of Cdc6p can drive both dephosphorylation and chromatin association of pol alpha. Overproduction of a mutant in which the N-terminus of Cdc6 has been deleted is unable to drive pol alpha chromatin binding. Since this mutant is still competent for pre-RC formation and DNA replication, we suggest that Cdc6p overproduction resets pol alpha chromatin binding by a mechanism which is independent of that used in pre-RC assembly.

MeSH Terms
CDC28 Protein Kinase, S cerevisiae/metabolism Cell Cycle Proteins/physiology Chromatin/metabolism Cyclin-Dependent Kinase Inhibitor Proteins DNA Polymerase I/metabolism DNA Primase/metabolism DNA Replication/physiology Fungal Proteins/genetics,physiology G1 Phase/physiology Mitosis/physiology Phosphorylation Recombinant Fusion Proteins Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins
Chemicals
CDC6 protein, S cerevisiae Cell Cycle Proteins Chromatin Cyclin-Dependent Kinase Inhibitor Proteins Fungal Proteins Recombinant Fusion Proteins SIC1 protein, S cerevisiae Saccharomyces cerevisiae Proteins CDC28 Protein Kinase, S cerevisiae DNA Primase DNA Polymerase I
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Desdouets C
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms EN6 3LD, UK.
Santocanale C
Drury L S
Perkins G
Foiani M
Plevani P
Diffley J F
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/17.14.4139
Published
1998-07-15
Pages
4139-46
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1170746
Subset
IM
Grants
Telethon · 1018 · Italy
External Links
PubMed source DOI: 10.1093/emboj/17.14.4139 PMC Full Text View journal details
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