...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular and Cellular Biology >DNA Length Dependence of the Single-Strand Annealing Pathway and the Role of Saccharomyces cerevisiae RAD59 in Double-Strand Break Repair
【24h】

DNA Length Dependence of the Single-Strand Annealing Pathway and the Role of Saccharomyces cerevisiae RAD59 in Double-Strand Break Repair

机译：单链退火途径的DNA长度依赖性和酿酒酵母RAD59在双链断裂修复中的作用

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

A DNA double-strand break (DSB) created by the HO endonuclease inSaccharomyces cerevisiae will stimulate recombination between flanking repeats by the single-strand annealing (SSA) pathway, producing a deletion. Previously the efficiency of SSA, using homologous sequences of different lengths, was measured in competition with that of a larger repeat further from the DSB, which ensured that nearly all cells would survive the DSB if the smaller region was not used (N. Sugawara and J. E. Haber, Mol. Cell. Biol. 12:563–575, 1992). Without competition, the efficiency with which homologous segments of 63 to 205 bp engaged in SSA was significantly increased. A sequence as small as 29 bp was used 0.2% of the time, and homology dependence was approximately linear up to 415 bp, at which size almost all cells survived. A mutant with a deletion of RAD59, a homologue of RAD52, was defective for SSA, especially when the homologous-sequence length was short; however, even with 1.17-kb substrates, SSA was reduced fourfold. DSB-induced gene conversion also showed a partial dependence on Rad59p, again being greatest when the homologous-sequence length was short. We found that Rad59p plays a role in removing nonhomologous sequences from the ends of single-stranded DNA when it invades a homologous DNA template, in a manner similar to that previously seen with srs2 mutants. Δrad59 affected DSB-induced gene conversion differently from msh3 and msh2, which are also defective in removing nonhomologous ends in both DSB-induced gene conversion and SSA. A msh3 rad59 double mutant was more severely defective in SSA than either single mutant.
机译：啤酒酵母中HO内切核酸酶产生的DNA双链断裂（DSB）将通过单链退火（SSA）途径刺激侧翼重复之间的重组，产生缺失。以前，使用不同长度的同源序列与距离DSB较大的重复序列竞争来测量SSA的效率，这确保了如果不使用较小的区域，几乎所有细胞都能在DSB中存活（N. Sugawara和JE Haber，分子细胞生物学杂志12：563-575，1992年）。在没有竞争的情况下，63至205 bp同源片段参与SSA的效率显着提高。 0.2％的时间使用的序列小至29 bp，并且同源性依赖性高达415 bp，呈线性关系，在此大小下，几乎所有细胞都可以存活。缺失 RAD59 （ RAD52 的同源物）的突变体具有SSA缺陷，尤其是当同源序列长度较短时;但是，即使使用1.17-kb的底物，SSA也减少了四倍。 DSB诱导的基因转化也显示出对Rad59p的部分依赖性，当同源序列长度较短时，也是最大的依赖性。我们发现Rad59p入侵同源DNA模板时，在从单链DNA末端去除非同源序列中起着作用，其作用方式与先前在 srs2 突变体中观察到的相似。 Δ rad59 对DSB诱导的基因转化的影响不同于 msh3 和 msh2 ，它们在去除DSB诱导的基因转化的非同源末端方面也存在缺陷和SSA。 msh3 rad59 双重突变体在SSA中的缺陷比任何一个单一突变体都严重。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号