Abstract
Replication factor C (RFC) is an essential, multi-subunit ATPase that functions in DNA replication, DNA repair, and DNA metabolism-related checkpoints. In order to investigate how the individual RFC subunits contribute to these functions in vivo, we undertook a genetic analysis of RFC genes from budding yeast. We isolated and characterized mutations in the RFC5 gene that could suppress the cold-sensitive phenotype of rfc1-1 mutants. Analysis of the RFC5 suppressors revealed that they could not suppress the elongated telomere phenotype, the sensitivity to DNA damaging agents, or the mutator phenotype of rfc1-1 mutants. Unlike the checkpoint-defective rfc5-1 mutation, the RFC5 suppressor mutations did not interfere with the methylmethane sulfonate- or hydroxyurea-induced phosphorylation of Rad53p. The Rfc5p suppressor substitutions mapped to amino acid positions in the conserved RFC box motifs IV-VII. Comparisons of the structures of related RFC box-containing proteins suggest that these RFC motifs may function to coordinate interactions between neighboring subunits of multi-subunit ATPases.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Alleles
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Amino Acid Motifs
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Amino Acid Sequence
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Cell Cycle Proteins / genetics*
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DNA Damage
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DNA Replication
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / genetics*
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Fungal Proteins / chemistry
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Fungal Proteins / genetics*
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Genes, Fungal*
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Homeodomain Proteins*
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Hydroxyurea / toxicity
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Methyl Methanesulfonate / toxicity
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Minor Histocompatibility Antigens
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Models, Molecular
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Molecular Sequence Data
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Mutation
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Phenotype
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Protein Structure, Quaternary
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Protein Subunits
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Proto-Oncogene Proteins c-bcl-2*
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Replication Protein C
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Repressor Proteins*
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Saccharomyces cerevisiae / drug effects
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins*
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Sequence Homology, Amino Acid
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Suppression, Genetic
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Telomere / genetics
Substances
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BCL2-related protein A1
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Cell Cycle Proteins
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DNA-Binding Proteins
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Fungal Proteins
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Homeodomain Proteins
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MATA1 protein, S cerevisiae
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Minor Histocompatibility Antigens
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Protein Subunits
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Proto-Oncogene Proteins c-bcl-2
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RFC1 protein, S cerevisiae
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Repressor Proteins
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Saccharomyces cerevisiae Proteins
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Methyl Methanesulfonate
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Replication Protein C
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Hydroxyurea