Fishman-Lobell J, Rudin N, Haber J E
Rosenstiel Center, Brandeis University, Boston, Massachusetts 02254-9110.
Mol Cell Biol. 1992 Mar;12(3):1292-303. doi: 10.1128/mcb.12.3.1292-1303.1992.
HO endonuclease-induced double-strand breaks in Saccharomyces cerevisiae can undergo recombination by two distinct and competing pathways. In a plasmid containing a direct repeat, in which one repeat is interrupted by an HO endonuclease cut site, gap repair yields gene conversions while single-strand annealing produces deletions. Consistent with predictions of the single-strand annealing mechanism, deletion formation is not accompanied by the formation of a reciprocal recombination product. Deletions are delayed 60 min when the distance separating the repeats is increased by 4.4 kb. Moreover, the rate of deletion formation corresponds to the time at which complementary regions become single stranded. Gap repair processes are independent of distance but are reduced in rad52 mutants and in G1-arrested cells.
酿酒酵母中 HO 核酸内切酶诱导的双链断裂可通过两种不同且相互竞争的途径进行重组。在含有同向重复序列的质粒中,其中一个重复序列被 HO 核酸内切酶切割位点打断,缺口修复产生基因转换,而单链退火产生缺失。与单链退火机制的预测一致,缺失形成不伴随相互重组产物的形成。当重复序列之间的距离增加 4.4 kb 时,缺失延迟 60 分钟形成。此外,缺失形成的速率与互补区域变为单链的时间相对应。缺口修复过程与距离无关,但在 rad52 突变体和 G1 期停滞的细胞中减少。