Aboussekhra A, Thoma F
Institut für Zellbiologie, Swiss Federal Institute of Technology (ETH)-Zürich, Hönggerberg, CH-8093 Zürich, Switzerland.
Genes Dev. 1998 Feb 1;12(3):411-21. doi: 10.1101/gad.12.3.411.
A high-resolution primer extension technique was used to study the relationships between repair, transcription, and mutagenesis in RNA polymerase III transcribed genes in Saccharomyces cerevisiae. The in vivo repair of UV-induced DNA damage by nucleotide excision repair (NER) and by photoreactivation is shown to be preferential for the nontranscribed strand (NTS) of the SNR6 gene. This is in contrast to RNA polymerase II genes in which the NER is preferential for the transcribed strand (TS). The repair-strand bias observed in SNR6 was abolished by inactivation of transcription in a snr6Delta2 mutant, showing a contribution of RNA polymerase III transcription in this phenomenon. The same strand bias for NER (slow in TS, fast in NTS) was discovered in the SUP4 gene, but only outside of the intragenic promoter element (box A). Unexpectedly, the repair in the transcribed box A was similar on both strands. The strand specificity as well as the repair heterogeneity determined in the transcribed strand of the SUP4 gene, correlate well with the previously reported site- and strand-specific mutagenesis in this gene. These findings present a novel view regarding the relationships between DNA repair, mutagenesis, and transcription.
采用高分辨率引物延伸技术研究酿酒酵母中RNA聚合酶III转录基因的修复、转录和诱变之间的关系。结果表明,核苷酸切除修复(NER)和光复活对紫外线诱导的DNA损伤的体内修复优先发生在SNR6基因的非转录链(NTS)上。这与RNA聚合酶II基因相反,在RNA聚合酶II基因中,NER优先发生在转录链(TS)上。在snr6Delta2突变体中,转录失活消除了在SNR6中观察到的修复链偏向,表明RNA聚合酶III转录在这一现象中起作用。在SUP4基因中也发现了相同的NER链偏向(TS中慢,NTS中快),但仅在基因内启动子元件(A框)之外。出乎意料的是,转录的A框中两条链上的修复相似。在SUP4基因转录链中确定的链特异性以及修复异质性,与该基因先前报道的位点和链特异性诱变密切相关。这些发现为DNA修复、诱变和转录之间的关系提供了新的观点。