The RAD7, RAD16, and RAD23 genes of Saccharomyces cerevisiae: Requirement for transcription-independent nucleotide excision repair in vitro and interactions between the gene products

Zhigang Wang, Shuguang Wei, Simon H. Reed, Xiaohua Wu, Jesper Q. Svejstrup, William J. Feaver, Roger D. Kornberg, Errol C. Friedberg

Research output: Contribution to journalArticle

65 Scopus citations

Abstract

Nucleotide excision repair (NER) is a biochemical process required for the repair of many different types of DNA lesions. In the yeast Saccharomyces cerevisiae, the RAD7, RAD16, and RAD23 genes have been specifically implicated in NER of certain transcriptionally repressed loci and in the nontranscribed strand of transcriptionally active genes. We have used a cell- free system to study the roles of the Rad7, Rad16, and Rad23 proteins in NER. Transcription-independent NER of a plasmid substrate was defective in rad7, rad16, and rad23 mutant extracts. Complementation studies with a previously purified NER protein complex (nucleotide excision repairosome) indicate that Rad23 is a component of the repairosome, whereas Rad7 and Rad16 proteins were not found in this complex. Complementation studies with rad4, rad7, rad16, and rad23 mutant extracts suggest physical interactions among these proteins. This conclusion was confirmed by experiments using the yeast two-hybrid assay, which demonstrated the following pairwise interactions: Rad4 with Rad23, Rad4 with Rad7, and Rad7 with Rad16. Additionally, interaction between the Rad7 and Rad16 proteins was demonstrated in vitro. Our results show that Rad7, Rad16, and Rad23 are required for transcription-independent NER in vitro. This process may involve a unique protein complex which is distinct from the repairosome and which contains at least the Rad4, Rad7, and Rad16 proteins.

Original languageEnglish (US)
Pages (from-to)635-643
Number of pages9
JournalMolecular and cellular biology
Volume17
Issue number2
DOIs
StatePublished - Feb 1997

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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