ORC and Cdc6p interact and determine the frequency of initiation of DNA replication in the genome

Liang, C., Weinreich, M., Stillman, B. (1995) ORC and Cdc6p interact and determine the frequency of initiation of DNA replication in the genome. Cell, 81 (5). pp. 667-76. ISSN 0092-8674 (Print)

URL: http://www.ncbi.nlm.nih.gov/pubmed/7774008
DOI: 10.1016/0092-8674(95)90528-6

Abstract

The origin recognition complex (ORC) binds replicators in the yeast S. cerevisiae in a manner consistent with it being an initiator protein for DNA replication. Two-dimensional (2D) gel techniques were used to examine directly initiation of chromosomal DNA replication in temperature-sensitive orc mutants. Unlike in wild-type cells, in orc2-1 and orc5-1 mutant cells, only a subset of replicators formed active origins of DNA replication at the permissive temperature. At the restrictive temperature, the number of active replicators was diminished further. Using a genetic screen, CDC6 was identified as a multicopy suppressor of orc5-1. 2D gel and biochemical analyses demonstrated that Cdc6p interacted functionally and physically with ORC. We suggest that ORC and Cdc6p form a prereplication complex at individual replicators and therefore cooperate to determine the frequency of initiation of DNA replication in the genome.

Item Type: Paper
Uncontrolled Keywords: Cell Cycle Proteins metabolism Chromosomes, Fungal Cloning Molecular DNA Replication DNA, Fungal biosynthesis DNA-Binding Proteins metabolism Electrophoresis, Gel, Two-Dimensional Genes, Lethal Genome Fungal Mutation Origin Recognition Complex Protein Binding Recombinant Fusion Proteins metabolism Replication Origin Repressor Proteins metabolism Saccharomyces cerevisiae genetics Saccharomyces cerevisiae Proteins Selection (Genetics) Suppression Genetic
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > DNA replication
CSHL Authors:
Communities: CSHL labs > Stillman lab
Highlight: Stillman, Bruce W.
Depositing User: CSHL Librarian
Date Deposited: 06 Mar 2012 15:09
Last Modified: 20 Jun 2017 19:45
Related URLs:
URI: http://repository.cshl.edu/id/eprint/25006

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