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Saccharomyces cerevisiae mms19 mutants are deficient in transcription-coupled and global nucleotide excision repair.

机译:酿酒酵母mms19突变体缺乏转录偶联和全局核苷酸切除修复。

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摘要

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.
机译:最近克隆的酿酒酵母MMS19基因似乎与核苷酸切除修复(NER)和转录都有关,NER /转录复合体TFIIH的成分也是如此。但是,与TFIIH不同,Mms19蛋白在高度纯化的体外系统中不会影响NER。为了研究Mms19在NER中的作用,我们分析了mms19破坏突变体的修复能力。我们发现该突变体的无细胞提取物在体外缺乏NER。由于mms19突变体仅对紫外线(UV)照射具有中等敏感性,因此此类突变体可能特别缺乏NER的两种模式之一,即转录偶联或整体基因组修复。为了研究这种可能性,我们分析了mms19突变体中核苷酸水平的环丁烷-嘧啶二聚体(CPD)的去除。该突变体的转录序列和非转录序列均无法检测到CPD的修复,这表明在转录偶联和整体基因组修复中都需要Mms19。我们的数据与其他人获得的数据相结合,表明Mms19是酵母中NER所必需的,尽管该蛋白似乎在此过程中起间接作用。

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