Role of yeast Rth1 nuclease and its homologs in mutation avoidance, DNA repair, and DNA replication

Robert E. Johnson, Gopala K. Kovvali, Louise Prakash, Satya Prakash

Research output: Contribution to journalArticle

49 Scopus citations

Abstract

The RTH1(RAD27) gene of Saccharomyces cerevisiae encodes a structure-specific endonuclease that cleaves 5'-ended single-stranded DNA at its junction with duplex DNA. Genetic and biochemical studies have indicated a role of Rth1 nuclease in the removal of RNA primers formed during DNA replication. The rth1Δ mutation confers temperature-sensitive lethality, and increases sensitivity to alkylating agents. The instability of repetitive DNA is greatly enhanced in the rth1Δ mutant. The conditional lethality of the rth1Δ mutation indicates that another nuclease can function in DNA replication in the absence of RTH1. RAD2, a homolog of RTH1, is required for nucleotide-excision repair. Here, we examine three other homologs of RTH1/RAD2 - YEN1, EXO1, and DIN7. Deletion of any of these genes in the rth1Δ strain has no effect on cell viability, suggesting the involvement of another, and as yet unidentified, nuclease in the maturation of Okazaki fragments. Our data also indicate that only RTH1 functions in the repair of alkylation damage. Deletions of YEN1, EXO1, DIN7, or RAD2, either singly or when combined with one another and with the rth1Δ mutation, have no effect on the rate of instability of dinucleotide repeats or on the rate of formation of large duplications in the CAN1 gene. These data provide evidence of a high degree of specificity for the role of RTH1 in DNA replication and in base-excision repair, and for the requirement of RAD2 in nucleotide-excision repair. The possibility that both Rth1 and Exo1 function in DNA mismatch repair is discussed.

Original languageEnglish (US)
Pages (from-to)21-29
Number of pages9
JournalCurrent Genetics
Volume34
Issue number1
DOIs
StatePublished - Jul 1 1998

Keywords

  • DNA replication
  • Exo1
  • Mismatch repair
  • Rth1
  • Yeast

ASJC Scopus subject areas

  • Genetics

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