Tu Y, Bates S, Pfeifer G P
Beckman Research Institute of the City of Hope, Department of Biology, Duarte, California 91010, USA.
J Biol Chem. 1997 Aug 15;272(33):20747-55. doi: 10.1074/jbc.272.33.20747.
Xeroderma pigmentosum (XP) and Cockayne syndrome (CS) cells have specific DNA repair defects. We had previously analyzed repair rates of cyclobutane pyrimidine dimers at nucleotide resolution along the human JUN gene in normal fibroblasts and found very efficient repair of sequences near the transcription initiation site but slow repair along the promoter. To investigate sequence-specific repair rate patterns in XP and CS cells, we conducted a similar analysis in XPA, XPB, XPC, XPD, and CSB fibroblasts. XPA cells were almost completely repair-deficient at all sequences analyzed. XPC cells repaired only the transcribed DNA strand beginning at position -20 relative to the transcription start site. Both XBP and XPD cells were deficient in repair of nontranscribed DNA and also very inefficiently repaired the transcribed strand including sequences near the transcription start site. CSB cells exhibited rapid repair near the transcription initiation site but were deficient in repair of sequences encountered by RNA polymerase during elongation (beginning at position +20). Since transcription of the JUN gene was UV-induced in all fibroblast strains, including CSB, the defective repair of the transcribed strand in CSB cannot be explained by a lack of transcription; rather, it appears to be a true DNA repair defect.
着色性干皮病(XP)和科凯恩综合征(CS)细胞具有特定的DNA修复缺陷。我们之前在正常成纤维细胞中,以核苷酸分辨率分析了人JUN基因上环丁烷嘧啶二聚体的修复率,发现转录起始位点附近的序列修复效率很高,但启动子区域的修复缓慢。为了研究XP和CS细胞中的序列特异性修复率模式,我们在XPA、XPB、XPC、XPD和CSB成纤维细胞中进行了类似分析。在所有分析的序列上,XPA细胞几乎完全缺乏修复能力。XPC细胞仅修复相对于转录起始位点从-20位置开始的转录DNA链。XBP和XPD细胞在非转录DNA的修复方面存在缺陷,并且对包括转录起始位点附近序列在内的转录链的修复效率也非常低。CSB细胞在转录起始位点附近表现出快速修复,但在RNA聚合酶延伸过程中遇到的序列(从+20位置开始)的修复方面存在缺陷。由于包括CSB在内的所有成纤维细胞系中JUN基因的转录都是紫外线诱导的,因此CSB中转录链的修复缺陷不能用缺乏转录来解释;相反,这似乎是一种真正的DNA修复缺陷。