Eukaryotic origin-dependent DNA replication in vitro reveals sequential action of DDK and S-CDK kinases.
Cell
; 146(1): 80-91, 2011 Jul 08.
Article
em En
| MEDLINE
| ID: mdl-21729781
ABSTRACT
Proper eukaryotic DNA replication requires temporal separation of helicase loading from helicase activation and replisome assembly. Using an in vitro assay for eukaryotic origin-dependent replication initiation, we investigated the control of these events. After helicase loading, we found that the Dbf4-dependent Cdc7 kinase (DDK) but not S phase cyclin-dependent kinase (S-CDK) is required for the initial origin recruitment of Sld3 and the Cdc45 helicase-activating protein. Likewise, in vivo, DDK drives early-firing-origin recruitment of Cdc45 before activation of S-CDK. After S-CDK activation, a second helicase-activating protein (GINS) and the remainder of the replisome are recruited to the origin. Finally, recruitment of lagging but not leading strand DNA polymerases depends on Mcm10 and DNA unwinding. Our studies identify distinct roles for DDK and S-CDK during helicase activation and support a model in which the leading strand DNA polymerase is recruited prior to origin DNA unwinding and RNA primer synthesis.
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Bases de dados:
MEDLINE
Assunto principal:
Saccharomyces cerevisiae
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Proteínas Serina-Treonina Quinases
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Quinases Ciclina-Dependentes
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Proteínas de Ciclo Celular
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Proteínas de Saccharomyces cerevisiae
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Replicação do DNA
Idioma:
En
Revista:
Cell
Ano de publicação:
2011
Tipo de documento:
Article