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RNA polymerase II transcription inhibits DNA repair by photolyase in the transcribed strand of active yeast genes.

机译:RNA聚合酶II转录通过活性酵母基因转录链中的光解酶抑制DNA修复。

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摘要

Yeast uses nucleotide excision repair (NER) and photolyase (photoreactivation) to repair cyclobutane pyrimidine dimers (CPDs) generated by ultraviolet light. In active genes, NER preferentially repairs the transcribed strand (TS). In contrast, we recently showed that photolyase preferentially repairs the non-transcribed strands (NTS) of the URA3 and HIS3 genes in minichromosomes. To test whether photoreactivation depends on transcription, repair of CPDs was investigated in the transcriptionally regulated GAL10 gene in a yeast strain deficient in NER [AMY3 (rad1Delta)]. In the active gene (cells grown in galactose), photoreactivation was fast in the NTS and slow in the TS demonstrating preferential repair of the NTS. In the inactive gene (cells grown in glucose), both strands were repaired at similar rates. This suggests that RNA polymerases II blocked at CPDs inhibit accessibility of CPDs to photolyase. In a strain in which both pathways are operational [W303-1a (RAD1)], no strand bias was observed either in the active or inactive gene, demonstrating that photoreactivation of the NTS compensates preferential repair of the TS by NER. Moreover, repair of the NTS was more quickly in the active gene than in the repressed gene indicating that transcription dependent disruption of chromatin facilitates repair of an active gene.
机译:酵母利用核苷酸切除修复(NER)和光裂解酶(光再活化)来修复紫外线产生的环丁烷嘧啶二聚体(CPD)。在活性基因中,NER优先修复转录的链(TS)。相反,我们最近发现,光解酶可优先修复微型染色体中URA3和HIS3基因的非转录链(NTS)。为了测试光激活是否取决于转录,研究了在NER [AMY3(rad1Delta)]缺陷型酵母菌株中,在转录调控的GAL10基因中修复了CPD。在活性基因(在半乳糖中生长的细胞)中,NTS中的光活化很快,而TS中的光活化很慢,这表明NTS的优先修复。在失活的基因(葡萄糖中生长的细胞)中,两条链都以相似的速率修复。这表明在CPDs处封闭的RNA聚合酶II抑制了CPDs对光裂解酶的可及性。在两种途径均起作用的菌株中[W303-1a(RAD1)],在活性或非活性基因中均未观察到链偏性,表明NTS的光再活化补偿了NER对TS的优先修复。此外,活性基因中NTS的修复比抑制基因中NTS的修复更快,这表明转录依赖性的染色质破坏促进了活性基因的修复。

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