Assembly of a complex containing Cdc45p, replication protein A, and Mcm2p at replication origins controlled by S-phase cyclin-dependent kinases and Cdc7p-Dbf4p kinase

Zou, L., Stillman, B. (2000) Assembly of a complex containing Cdc45p, replication protein A, and Mcm2p at replication origins controlled by S-phase cyclin-dependent kinases and Cdc7p-Dbf4p kinase. Molecular and Cellular Biology, 20 (9). pp. 3086-3096. ISSN 0270-7306

URL: http://www.ncbi.nlm.nih.gov/pubmed/10757793
DOI: 10.1128/​MCB.20.9.3086-3096.2000

Abstract

In Saccharomyces cerevisiae, replication origins are activated with characteristic timing during S phase. S-phase cyclin-dependent kinases (S-CDKs) and Cdc7p-Dbf4p kinase are required far origin activation throughout S phase. The activation of S-CDKs leads to association of Cdc45p with chromatin, raising the possibility that Cdc45p defines the assembly of a new complex at each origin. Here we show that bath Cdc45p and replication protein A (RPA) bind to Mcm2p at the G(1)-S transition in an S-CDK-dependent manner. During S phase, Cdc45p associates with different replication origins at specific times, The origin associations of Cdc45p and RPA are mutually dependent, and both S-CDKs and Cdc7p-Dbf4p are required for efficient binding of Cdc45p to origins. These findings suggest that S-CDKs and Cdc7p-Dbf4p promote loading of Cdc45p and RPA onto a preformed prereplication complex at each origin with preprogrammed timing. The ARS1 association of Mcm2p, but not that of the origin recognition complex, is diminished by disruption of the B2 element of ARS1, a potential origin DNA-unwinding element. Cdc45p is required for recruiting DNA polymerase alpha onto chromatin, and it associates with Mcm2p, RPA, and DNA polymerase epsilon only during S phase, These results suggest that the complex containing Cdc45p, RPA, and MCMs is involved in origin unwinding and assembly of replication forks at each origin.

Item Type: Paper
Uncontrolled Keywords: CHROMOSOMAL DNA-REPLICATION chromosomal DNA replicatin chrom SACCHAROMYCES-CEREVISIAE Saccharomyces Cerevisiae CELL-CYCLE cell cycle BUDDING YEAST budding yeast MINICHROMOSOME MAINTENANCE minichromosome maintenance RECOGNITION COMPLEX recognition complex EUKARYOTIC CELLS eukaryotic cells FISSION YEAST fission yeast CDC6 PROTEIN CDC6 protein cdc6 ARS ELEMENTS ARS elements
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
organism description > yeast > Saccharomyces
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date Deposited: 17 Feb 2012 16:04
Last Modified: 20 Jun 2017 18:52
PMCID: PMC85601
Related URLs:
URI: http://repository.cshl.edu/id/eprint/25152

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