Morse Nerys R, Meniel Valerie, Waters Raymond
School of Biological Sciences, University of Wales Swansea, Singleton Park, Swansea SA2 8PP, UK.
Nucleic Acids Res. 2002 Apr 15;30(8):1799-807. doi: 10.1093/nar/30.8.1799.
Photoreactive repair (PR) of cyclobutane pyrimidine dimers (CPDs) was mapped at nucleotide resolution in nucleotide excision repair (NER) proficient and deficient strains for the transcriptionally active and inactive MFA2 gene. Enhanced PR in the control region occurred in areas where no nucleosomes were present, particularly linker regions in the alpha mating type. The presence of excision plus transcriptional activation alleviated this preference, causing repair in the regions that were linker and core in the alpha mating type to be the same in this strain. Transcription had no effect on photoreactive repair in transcribed and downstream regions of MFA2, where similar rates were observed for specific CPDs in both strands. The presence of positioned nucleosomes in alpha mating types revealed slow repair in the nucleosome core, with faster repair occurring at the 3' and 5' edges. These data support the evidence that CPDs are repaired quicker in nucleosome-free regions and at edges of nucleosomes. CPDs in the linker regions are repaired more efficiently in the transcriptionally inactive strains, suggesting that nucleosome movement associated with transcription of MFA2 hampers PR irrespective of the strand. Proficient NER influenced PR in the TATA and Mcm1 binding sites by enhancing it, particularly when transcription was activated.
在转录活性和非活性的MFA2基因中,以核苷酸分辨率绘制了环丁烷嘧啶二聚体(CPD)的光反应修复(PR)图谱,该图谱针对核苷酸切除修复(NER)能力正常和缺陷的菌株。对照区域中增强的PR发生在不存在核小体的区域,特别是在α交配型中的连接区。切除加转录激活的存在减轻了这种偏好,导致该菌株中α交配型中连接区和核心区的修复情况相同。转录对MFA2转录区和下游区域的光反应修复没有影响,在这两个区域中,两条链上特定CPD的修复速率相似。α交配型中定位核小体的存在表明核小体核心区域的修复较慢,而在3'和5'边缘修复较快。这些数据支持了CPD在无核小体区域和核小体边缘修复更快的证据。连接区中的CPD在转录非活性菌株中修复更有效,这表明与MFA2转录相关的核小体移动会阻碍PR,而与链无关。熟练的NER通过增强PR影响TATA和Mcm1结合位点的PR,特别是在转录激活时。