Moore J K, Haber J E
Rosenstiel Center and Department of Biology, Brandeis University, Waltham, Massachusetts 02254-9110, USA.
Nature. 1996 Oct 17;383(6601):644-6. doi: 10.1038/383644a0.
Non-homologous repair of broken chromosomes in Saccharomyces cerevisiae can be studied at a defined location by expressing the site-specific HO endonuclease that cuts the mating-type (MAT) locus. When homologous recombination is prevented, most double-strand breaks are repaired by non-homologous end-joinings similar to those observed in mammalian cells. About 1% of non-homologous repair events were exceptional, having 'captured' approximately 100 base pairs of DNA within the HO cleavage site. In each case, the insertion came from yeast's retrotransposon Tyl element. Four of the five contained the R-U5 region, which is the first part of Tyl messenger RNA to be converted to complementary DNA. The capture of cDNA fragments at the sites of double-strand breaks may account for the way that pseudogenes and long and short interspersed sequences (LINES and SINES) have been inserted at many locations in the mammalian genome.
通过表达切割交配型(MAT)位点的位点特异性HO内切核酸酶,可以在特定位置研究酿酒酵母中破碎染色体的非同源修复。当同源重组被阻止时,大多数双链断裂通过类似于在哺乳动物细胞中观察到的非同源末端连接进行修复。约1%的非同源修复事件是例外情况,在HO切割位点内“捕获”了大约100个碱基对的DNA。在每种情况下,插入片段都来自酵母的逆转座子Tyl元件。五个中有四个包含R-U5区域,这是Tyl信使RNA转化为互补DNA的第一部分。双链断裂位点处cDNA片段的捕获可能解释了假基因以及长散在序列和短散在序列(LINEs和SINEs)在哺乳动物基因组中许多位置插入的方式。