The N-degron protein degradation strategy for investigating the function of essential genes

requirement for replication protein A and proliferating cell nuclear antigen proteins for nucleotide excision repair in yeast extracts

Wen-Ya Huang, William J. Feaver, Alan E. Tomkinson, Errol C. Friedberg

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Nucleotide excision repair (NER) of DNA in the yeast Saccharomyces cerevisiae and in human cells has been shown to be a biochemically complex process involving multiple gene products. In yeast, the involvement of the DNA replication accessory proteins, replication protein A (RPA1) and proliferating cell nuclear antigen (PCNA) in NER has not been demonstrated genetically. In this study we have generated temperature-degradable rfa1 and pcna mutants and show that these mutants are defective in NER in vitro under conditions that promote degradation of the RFA1 and PCNA gene products. We also demonstrate a physical interaction between RPA1 protein and subunits of the RNA polymerase II basal transcription factor IIH (TFIIH). Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)183-194
Number of pages12
JournalMutation Research - DNA Repair
Volume408
Issue number3
DOIs
Publication statusPublished - 1998 Sep 11

Fingerprint

Replication Protein A
Essential Genes
Proliferating Cell Nuclear Antigen
Nuclear Proteins
DNA Repair
Yeast
Proteolysis
Repair
Nucleotides
Genes
Yeasts
Degradation
Transcription Factor TFIIH
Proteins
RNA Polymerase II
DNA
Protein Subunits
Accessories
DNA Replication
Saccharomyces cerevisiae

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Toxicology
  • Genetics

Cite this

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abstract = "Nucleotide excision repair (NER) of DNA in the yeast Saccharomyces cerevisiae and in human cells has been shown to be a biochemically complex process involving multiple gene products. In yeast, the involvement of the DNA replication accessory proteins, replication protein A (RPA1) and proliferating cell nuclear antigen (PCNA) in NER has not been demonstrated genetically. In this study we have generated temperature-degradable rfa1 and pcna mutants and show that these mutants are defective in NER in vitro under conditions that promote degradation of the RFA1 and PCNA gene products. We also demonstrate a physical interaction between RPA1 protein and subunits of the RNA polymerase II basal transcription factor IIH (TFIIH). Copyright (C) 1998 Elsevier Science B.V.",
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AU - Tomkinson, Alan E.

AU - Friedberg, Errol C.

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