Liang, C., Weinreich, M., Stillman, B. (June 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)
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: | 2 June 1995 |
| Date Deposited: | 06 Mar 2012 15:09 |
| Last Modified: | 20 Jun 2017 19:45 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/25006 |
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