ORC and Cdc6p interact and determine the frequency of initiation of DNA replication in the genome.
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.
Mesh Terms:
Cell Cycle Proteins, Chromosomes, Fungal, Cloning, Molecular, DNA Replication, DNA, Fungal, DNA-Binding Proteins, Electrophoresis, Gel, Two-Dimensional, Genes, Lethal, Genome, Fungal, Mutation, Origin Recognition Complex, Protein Binding, Recombinant Fusion Proteins, Replication Origin, Repressor Proteins, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Selection, Genetic, Suppression, Genetic
Cell Cycle Proteins, Chromosomes, Fungal, Cloning, Molecular, DNA Replication, DNA, Fungal, DNA-Binding Proteins, Electrophoresis, Gel, Two-Dimensional, Genes, Lethal, Genome, Fungal, Mutation, Origin Recognition Complex, Protein Binding, Recombinant Fusion Proteins, Replication Origin, Repressor Proteins, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Selection, Genetic, Suppression, Genetic
Cell
Date: Jun. 02, 1995
PubMed ID: 7774008
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