Cyclin B-cdk1 kinase stimulates ORC- and Cdc6-independent steps of semiconservative plasmid replication in yeast nuclear extracts.

Détails

ID Serval
serval:BIB_12091
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Cyclin B-cdk1 kinase stimulates ORC- and Cdc6-independent steps of semiconservative plasmid replication in yeast nuclear extracts.
Périodique
Molecular and Cellular Biology
Auteur(s)
Duncker B.P., Pasero P., Braguglia D., Heun P., Weinreich M., Gasser S.M.
ISSN
0270-7306
Statut éditorial
Publié
Date de publication
1999
Volume
19
Numéro
2
Pages
1226-1241
Langue
anglais
Notes
Publication types: In Vitro ; Journal Article
Résumé
Nuclear extracts from Saccharomyces cerevisiae cells synchronized in S phase support the semiconservative replication of supercoiled plasmids in vitro. We examined the dependence of this reaction on the prereplicative complex that assembles at yeast origins and on S-phase kinases that trigger initiation in vivo. We found that replication in nuclear extracts initiates independently of the origin recognition complex (ORC), Cdc6p, and an autonomously replicating sequence (ARS) consensus. Nonetheless, quantitative density gradient analysis showed that S- and M-phase nuclear extracts consistently promote semiconservative DNA replication more efficiently than G1-phase extracts. The observed semiconservative replication is compromised in S-phase nuclear extracts deficient for the Cdk1 kinase (Cdc28p) but not in extracts deficient for the Cdc7p kinase. In a cdc4-1 G1-phase extract, which accumulates high levels of the specific Clb-Cdk1 inhibitor p40(SIC1), very low levels of semiconservative DNA replication were detected. Recombinant Clb5-Cdc28 restores replication in a cdc28-4 S-phase extract yet fails to do so in the cdc4-1 G1-phase extract. In contrast, the addition of recombinant Xenopus CycB-Cdc2, which is not sensitive to inhibition by p40(SIC1), restores efficient replication to both extracts. Our results suggest that in addition to its well-characterized role in regulating the origin-specific prereplication complex, the Clb-Cdk1 complex modulates the efficiency of the replication machinery itself.
Mots-clé
Animals, CDC2 Protein Kinase/genetics, CDC2 Protein Kinase/metabolism, Cell Cycle Proteins/genetics, Cell Cycle Proteins/metabolism, Cell Nucleus/metabolism, Cyclin B/genetics, Cyclin B/metabolism, DNA Replication/genetics, DNA, Fungal/biosynthesis, DNA, Fungal/genetics, DNA-Binding Proteins/genetics, DNA-Binding Proteins/metabolism, Fungal Proteins/genetics, Fungal Proteins/metabolism, G1 Phase, Mutation, Origin Recognition Complex, Plasmids/genetics, Plasmids/metabolism, Recombinant Proteins/genetics, Recombinant Proteins/metabolism, S Phase, Saccharomyces cerevisiae/genetics, Saccharomyces cerevisiae/metabolism, Saccharomyces cerevisiae Proteins, Xenopus
Pubmed
Web of science
Création de la notice
19/11/2007 13:02
Dernière modification de la notice
20/08/2019 13:39
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