Rusyn Ivan, Fry Rebecca C, Begley Thomas J, Klapacz Joanna, Svensson J Peter, Ambrose Mark, Samson Leona D
Center for Environmental Health Sciences and Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America.
PLoS One. 2007 Nov 28;2(11):e1252. doi: 10.1371/journal.pone.0001252.
Upon exposure to DNA damaging agents, Saccharomyces cerevisiae respond by activating a massive transcriptional program that reflects the fact that "DNA damaging" agents also damage other cellular macromolecules. To identify the transcriptional response that is specific to DNA damage, we have modulated the first two enzymes in the base excision repair (BER) pathway generating yeast strains with varied levels of the repair intermediates, abasic sites or strand breaks. We show that the number of abasic sites is significantly increased when the 3-methyladenine DNA glycosylase (Mag): AP endonuclease (Apn1) ratio is increased and that spontaneous frame shift mutation is considerably elevated when either Mag, or Mag plus Apn1, expression is elevated. Expression profiling identified 633 ORFs with differential expression associated with BER modulation. Analysis of transcriptional networks associated with the accumulation of DNA repair intermediates identifies an enrichment for numerous biological processes. Moreover, most of the BER-activated transcriptional response was independent of the classical yeast environmental stress response (ESR). This study highlights that DNA damage in the form of abasic sites or strand breaks resulting from BER modulation is a trigger for substantial genome-wide change and that this response is largely ESR-independent. Taken together, these results suggest that a branch point exists in the current model for DNA damage-signaled transcription in S. cerevisiae.
在暴露于DNA损伤剂时,酿酒酵母通过激活一个大规模的转录程序做出反应,这反映了“DNA损伤”剂也会损伤其他细胞大分子这一事实。为了鉴定对DNA损伤特异的转录反应,我们调节了碱基切除修复(BER)途径中的前两种酶,从而产生了具有不同水平修复中间体、无碱基位点或链断裂的酵母菌株。我们发现,当3-甲基腺嘌呤DNA糖基化酶(Mag):AP核酸内切酶(Apn1)的比例增加时,无碱基位点的数量显著增加;当Mag或Mag加Apn1的表达升高时,自发移码突变也大幅增加。表达谱分析鉴定出633个与BER调节相关的差异表达开放阅读框(ORF)。对与DNA修复中间体积累相关的转录网络的分析确定了许多生物过程的富集。此外,大多数BER激活的转录反应独立于经典的酵母环境应激反应(ESR)。这项研究突出表明,由BER调节导致的无碱基位点或链断裂形式的DNA损伤是全基因组发生实质性变化的触发因素,并且这种反应在很大程度上独立于ESR。综上所述,这些结果表明,在当前酿酒酵母DNA损伤信号转录模型中存在一个分支点。