Saccharomyces cerevisiae mms19 mutants are deficient in transcription-coupled and global nucleotide excision repair.
The recently cloned Saccharomyces cerevisiae MMS19 gene appears to be involved in both nucleotide excision repair (NER) and transcription, which is also the case for components of the NER/transcription complex TFIIH. Unlike TFIIH however, the Mms19 protein does not affect NER in a highly purified in vitro system. In order to investigate the role of Mms19 in NER, we have analysed the repair capacity of the mms19 disruption mutant. We find that a cell-free extract of this mutant is deficient for NER in vitro. Since mms19 mutants are only moderately sensitive to irradiation with ultraviolet (UV) light, it is possible that such mutants are specifically deficient in one of the two modes of NER, i.e. transcription-coupled or global genome repair. To investigate this possibility, we have analysed the removal of cyclobutane-pyrimidine dimers (CPDs) at the nucleotide level in an mms19 mutant. Repair of CPDs was not detectable for both transcribed and non-transcribed sequences in this mutant, demonstrating a requirement for Mms19 in both transcription-coupled and global genome repair. Our data, combined with those obtained by others, suggest that Mms19 is required for NER in yeast, although it seems likely that the protein plays an indirect role in this process.
Lombaerts
M
M
Laboratory of Molecular Genetics, Leiden Institute of Chemistry, Leiden University, PO Box 9502, 2300 RA Leiden, The Netherlands.
Tijsterman
M
M
Verhage
R A
RA
Brouwer
J
J
D004260:DNA Repair / Q000235:genetics*; D004271:DNA, Fungal / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics*; D007700:Kinetics; D009154:Mutation; D009711:Nucleotides / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011740:Pyrimidine Dimers / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D014158:Transcription, Genetic / Q000235:genetics*; D014466:Ultraviolet Rays
D004271:DNA, Fungal; D005656:Fungal Proteins; C103317:MET18 protein, S cerevisiae; D009711:Nucleotides; D011740:Pyrimidine Dimers; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors