A genomics-based screen for yeast mutants with an altered recombination/end-joining repair ratio.
A genomics-based screen for yeast mutants with an altered recombination/end-joining repair ratio.
复制标题
基于基因组学的酵母突变体筛选，具有改变的重组/末端连接修复比率。
DOI：
10.1093/genetics/162.2.677
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发表时间：
2002
期刊：
影响因子：
3.3
通讯作者：
Wilson,ThomasE
中科院分区：
文献类型：
--
作者：
Wilson,ThomasE
We recently described a yeast assay suitable for genetic screening in which simple religation nonhomologous end-joining (NHEJ) and single-strand annealing (SSA) compete for repair of an I-SceI-created double-strand break. Here, the required allele has been introduced into an array of 4781MATadeletion mutants and each strain screened individually. Two mutants (rad52andsrs2) showed a clear increase in the NHEJ/SSA ratio due to preferential impairment of SSA, but no mutant increased the absolute frequency of NHEJ significantly above the wild-type level. Seven mutants showed a decreased NHEJ/SSA ratio due to frank loss of NHEJ, which corresponded to all known structural/catalytic NHEJ components (yku70, yku80, dnl4, lif1, rad50, mre11, andxrs2); no new mutants in this category were identified. A clearly separable and surprisingly large set of 16 other mutants showed partial defects in NHEJ. Further examination of these revealed thatNEJ1can entirely account for the mating-type regulation of NHEJ, but that this regulatory role was distinct from the postdiauxic/stationary-phase induction of NHEJ that was deficient in other mutants (especiallydoa1, fyv6, andmck1). These results are discussed in the context of the minimal set of required proteins and regulatory inputs for NHEJ.
