Dominant mutations in three different subunits of replication factor C suppress replication defects in yeast PCNA mutants
| Autor: | Neelam S. Amin, K M Tuffo, Connie Holm |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1999 |
| Předmět: |
DNA Replication
Saccharomyces cerevisiae Proteins Mutant Saccharomyces cerevisiae Biology medicine.disease_cause DNA polymerase delta RFC2 Fungal Proteins Minor Histocompatibility Antigens Replication factor C Proliferating Cell Nuclear Antigen Genetics medicine Humans Replication Protein C Genes Suppressor Alleles Genes Dominant Homeodomain Proteins Mutation DNA replication Processivity Molecular biology Proliferating cell nuclear antigen DNA-Binding Proteins Repressor Proteins Proto-Oncogene Proteins c-bcl-2 biology.protein Research Article |
| Popis: | To identify proteins that interact with the yeast proliferating cell nuclear antigen (PCNA), we used a genetic approach to isolate mutations that compensate for the defects in cold-sensitive (Cs−) mutants of yeast PCNA (POL30). Because the cocrystal structure of human PCNA and a p21WAF1/CIP1 peptide shows that the interdomain region of PCNA is a site of p21 interaction, we specifically looked for new mutations that suppress mutations in the equivalent region of yeast PCNA. In independent screens using three different Cs− mutants, we identified spontaneously arising dominant suppressor mutations in the RFC3 gene. In addition, dominant suppressor mutations were identified in the RFC1 and RFC2 genes using a single pol30 mutant. An intimate association between PCNA and RFC1p, RFC2p, and RFC3p is suggested by the allele-restricted suppression of 10 different pol30 alleles by the RFC suppressors. RFC1, RFC2, and RFC3 encode three of the five subunits of the replication factor C complex, which is required to load PCNA onto DNA in reconstituted DNA replication reactions. Genomic sequencing reveals a common region in RFC1p, RFC2p, and RFC3p that is important for the functional interaction with PCNA. Biochemical analysis of the wild type and mutant PCNA and RFC3 proteins shows that mutant RFC3p enhances the production of long DNA products in pol δ-dependent DNA synthesis, which is consistent with an increase in processivity. |
| Databáze: | OpenAIRE |
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