Prado F, González-Barrera S, Aguilera A
Departamento de Genética, Facultad de Biología, Universidad de Sevilla, Spain.
Mol Gen Genet. 2000 Feb;263(1):73-80. doi: 10.1007/pl00008677.
We have devised a system for isolating yeast DNA sequences that are able to act as initiators of recombination leading to deletions in mitotically growing yeast cells. This system has allowed us to identify the FRT site of the 2-micron site-specific recombinase Flp as such a sequence. We show that Flp is able to initiate recombination leading to deletions at a single FRT site in cir(o) strains. These results indicate that Flp is able to cleave a single FRT site, supporting the observation that the mechanism of cleavage by Flp is trans-horizontal. Interestingly, Flp can induce homologous recombination in both a RAD52-dependent and RAD52-independent manner. Our work provides a new system for the study of homologous recombination leading to deletions, in which the initiation step can be efficiently controlled. We discuss the possibility that Flp-induced, RAD52-independent events occur by single-strand annealing.
我们设计了一种系统，用于分离能够作为重组起始位点、导致有丝分裂生长的酵母细胞发生缺失的酵母DNA序列。该系统使我们能够鉴定出2μm位点特异性重组酶Flp的FRT位点就是这样一种序列。我们表明，Flp能够在cir(o)菌株的单个FRT位点起始导致缺失的重组。这些结果表明Flp能够切割单个FRT位点，支持了Flp的切割机制是反式水平的这一观察结果。有趣的是，Flp可以以RAD52依赖和RAD52非依赖的方式诱导同源重组。我们的工作为研究导致缺失的同源重组提供了一个新系统，其中起始步骤可以得到有效控制。我们讨论了Flp诱导的、RAD52非依赖事件通过单链退火发生的可能性。
