Abstract
This report describes the isolation of ORC5, the gene encoding the fifth largest subunit of the origin recognition complex, and the properties of mutants with a defective allele of ORC5. The orc5-1 mutation caused temperature-sensitive growth and, at the restrictive temperature, caused cell cycle arrest. At the permissive temperature, the orc5-1 mutation caused an elevated plasmid loss rate that could be suppressed by additional tandem origins of DNA replication. The sequence of ORC5 revealed a potential ATP binding site, making Orc5p a candidate for a subunit that mediates the ATP-dependent binding of ORC to origins. Genetic interactions among orc2-1 and orc5-1 and other cell cycle genes provided further evidence for a role for the origin recognition complex (ORC) in DNA replication. The silencing defect caused by orc5-1 strengthened previous connections between ORC and silencing, and combined with the phenotypes caused by orc2 mutations, suggested that the complex itself functions in both processes.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Cell Cycle / genetics
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Cell Cycle Proteins / metabolism
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Cloning, Molecular
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Crosses, Genetic
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DNA Replication*
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DNA, Fungal / metabolism
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / genetics*
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DNA-Binding Proteins / metabolism
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DNA-Binding Proteins / physiology
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Genes, Fungal*
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Molecular Sequence Data
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Mutation
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Origin Recognition Complex
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Phenotype
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Plasmids / biosynthesis
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Polymerase Chain Reaction
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Repressor Proteins / chemistry
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Repressor Proteins / genetics*
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Repressor Proteins / metabolism
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Repressor Proteins / physiology
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Saccharomyces cerevisiae / cytology
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae / growth & development
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins
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Temperature
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Time Factors
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Transcription, Genetic
Substances
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Cell Cycle Proteins
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DNA, Fungal
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DNA-Binding Proteins
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ORC2 protein, S cerevisiae
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ORC5 protein, S cerevisiae
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Origin Recognition Complex
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Repressor Proteins
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Saccharomyces cerevisiae Proteins
Associated data
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GENBANK/U24187
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GENBANK/U34860
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GENBANK/U34861
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GENBANK/U34862