Tsubouchi H, Ogawa H
Department of Biology, Graduate School of Science, Osaka University, Osaka 560-0043, Japan.
Mol Biol Cell. 2000 Jul;11(7):2221-33. doi: 10.1091/mbc.11.7.2221.
The MRE11, RAD50, and XRS2 genes of Saccharomyces cerevisiae are involved in the repair of DNA double-strand breaks (DSBs) produced by ionizing radiation and by radiomimetic chemicals such as methyl methanesulfonate (MMS). In these mutants, single-strand DNA degradation in a 5' to 3' direction from DSB ends is reduced. Multiple copies of the EXO1 gene, encoding a 5' to 3' double-strand DNA exonuclease, were found to suppress the high MMS sensitivity of these mutants. The exo1 single mutant shows weak MMS sensitivity. When an exo1 mutation is combined with an mre11 mutation, both repair of MMS-induced damage and processing of DSBs are more severely reduced than in either single mutant, suggesting that Exo1 and Mre11 function independently in DSB processing. During meiosis, transcription of the EXO1 gene is highly induced. In meiotic cells, the exo1 mutation reduces the processing of DSBs and the frequency of crossing over, but not the frequency of gene conversion. These results suggest that Exo1 functions in the processing of DSB ends and in meiotic crossing over.
酿酒酵母的MRE11、RAD50和XRS2基因参与由电离辐射以及诸如甲磺酸甲酯(MMS)等拟辐射化学物质产生的DNA双链断裂(DSB)的修复。在这些突变体中,从DSB末端向5'至3'方向的单链DNA降解减少。发现编码5'至3'双链DNA外切核酸酶的EXO1基因的多个拷贝可抑制这些突变体对MMS的高敏感性。exo1单突变体表现出较弱的MMS敏感性。当exo1突变与mre11突变结合时,MMS诱导损伤的修复和DSB的加工均比任一单突变体更严重地减少,这表明Exo1和Mre11在DSB加工中独立发挥作用。在减数分裂期间,EXO1基因的转录被高度诱导。在减数分裂细胞中,exo1突变减少了DSB的加工和交叉频率,但不影响基因转换频率。这些结果表明Exo1在DSB末端加工和减数分裂交叉中发挥作用。