Parket A, Inbar O, Kupiec M
Department of Molecular Microbiology and Biotechnology, Tel Aviv University, Ramat Aviv, Israel.
Genetics. 1995 May;140(1):67-77. doi: 10.1093/genetics/140.1.67.
The Ty retrotransposons are the main family of dispersed repeated sequences in the yeast Saccharomyces cerevisiae. These elements are flanked by a pair of long terminal direct repeats (LTRs). Previous experiments have shown that Ty elements recombine at low frequencies, despite the fact that they are present in 30 copies per genome. This frequency is not highly increased by treatments that cause DNA damage, such as UV irradiation. In this study, we show that it is possible to increase the recombination level of a genetically marked Ty by creating a double-strand break in it. This break is repaired by two competing mechanisms: one of them leaves a single LTR in place of the Ty, and the other is a gene conversion event in which the marked Ty is replaced by an ectopically located one. In a strain in which the marked Ty has only one LTR, the double-strand break is repaired by conversion. We have also measured the efficiency of repair and monitored the progression of the cells through the cell-cycle. We found that in the presence of a double-strand break in the marked Ty, a proportion of the cells is unable to resume growth.
Ty逆转座子是酿酒酵母中分散重复序列的主要家族。这些元件两侧是一对长末端正向重复序列(LTRs)。先前的实验表明,尽管每个基因组中存在30个拷贝,但Ty元件的重组频率很低。通过诸如紫外线照射等导致DNA损伤的处理,这种频率并没有大幅增加。在本研究中,我们表明通过在基因标记的Ty中产生双链断裂,可以提高其重组水平。这种断裂通过两种相互竞争的机制进行修复:其中一种机制是留下一个单一的LTR来取代Ty,另一种是基因转换事件,其中标记的Ty被异位的Ty所取代。在标记的Ty只有一个LTR的菌株中,双链断裂通过转换进行修复。我们还测量了修复效率,并监测了细胞在细胞周期中的进程。我们发现,在标记的Ty中存在双链断裂的情况下,一部分细胞无法恢复生长。