RAD59 and RAD1 cooperate in translocation formation by single-strand annealing in Saccharomyces cerevisiae.
RAD59 and RAD1 cooperate in translocation formation by single-strand annealing in Saccharomyces cerevisiae.
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DOI：
10.1007/s00294-009-0282-6
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发表时间：
2010-02
期刊：
影响因子：
2.5
通讯作者：
Bailis, Adam M.
中科院分区：
文献类型：
--
作者：
Pannunzio, Nicholas R.;Manthey, Glenn M.;Bailis, Adam M.
关键词：
对萌芽酵母酿酒酵母的研究表明，急性辐射后双链断裂修复的相当大一部分涉及重复基因组元件之间的同源重组。我们之前已经描述了一种在酿酒酵母中的实验，它允许我们建模多个断裂的修复如何通过单链退火(SSA)导致染色体易位的形成，并发现Rad59，一个单链DNA退火蛋白Rad52的并列，在这一过程中至关重要。我们已经构建了几个rad59错义等位基因，以更深入地研究其功能。对这些突变体的鉴定显示，易位形成和自发的直接重复重组都存在比例缺陷，这也被认为是由SSA引起的。将rad59错义等位基因与Rad1的零等位基因相结合，基本上抑制了在Rad1缺失的单个突变体中观察到的低频率易位。Rad1的零等位基因编码了从退火中间体中移除非同源尾巴所需的核酸酶的一个亚单位。这些数据表明，Rad59在SSA易位形成中至少有一个作用是支持切割非同源尾巴所需的机制。
Studies in the budding yeast, Saccharomyces cerevisiae, have demonstrated that a substantial fraction of double-strand break repair following acute radiation exposure involves homologous recombination between repetitive genomic elements. We have previously described an assay in S. cerevisiae that allows us to model how repair of multiple breaks leads to the formation of chromosomal translocations by single-strand annealing (SSA) and found that Rad59, a paralog of the single-stranded DNA annealing protein Rad52, is critically important in this process. We have constructed several rad59 missense alleles to study its function more closely. Characterization of these mutants revealed proportional defects in both translocation formation and spontaneous direct-repeat recombination, which is also thought to occur by SSA. Combining the rad59 missense alleles with a null allele of RAD1, which encodes a subunit of a nuclease required for the removal of non-homologous tails from annealed intermediates, substantially suppressed the low frequency of translocations observed in rad1-null single mutants. These data suggest that at least one role of Rad59 in translocation formation by SSA is supporting the machinery required for cleavage of non-homologous tails.
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