4.3 Article

The global role for Cdc13 and Yku70 in preventing telomere resection across the genome

期刊

DNA REPAIR
卷 62, 期 -, 页码 8-17

出版社

ELSEVIER SCIENCE BV
DOI: 10.1016/j.dnarep.2017.11.010

关键词

Telomere; Resection; Yku; Rad52; Yeast; Foci

资金

  1. Intramural Research Program of the National Institute of Environmental Health Sciences, NIH [1 Z01 ES065073]
  2. National Institutes of Health [ES024872]
  3. American Cancer Society [129182-RSG-16-043-01-DMC]

向作者/读者索取更多资源

Yeast Cdc13 protein (related to human CTC1) maintains telomere stability by preventing 5'-3' end resection. While Cdc13 and Ylcu70/Yku80 proteins appear to prevent excessive resection, their combined contribution to maintenance of telomere ends across the genome and their relative roles at specific ends of different chromosomes have not been addressable because Cdc13 and Yku70/Yku80 double mutants are sickly. Using our PFGE-shift approach where large resected molecules have slower pulse field gel electrophoresis mobilities, along with methods for maintaining viable double mutants, we address end-resection on most chromosomes as well as telomere end differences. In this global approach to looking at ends of most chromosomes, we identify chromosomes with 1-end resections and end-preferences. We also identify chromosomes with resection at both ends, previously not possible. 10-20% of chromosomes exhibit PFGE-shift when cdc13-1 cells are switched to restrictive temperature (37 degrees C). In yku70 Delta cdc13-1 mutants, there is a telomere resection storm with approximately half the chromosomes experiencing at least 1-end resection, similar to 10 kb/telomere, due to exonucleasel and many exhibiting 2-end resection. Unlike for random internal chromosome breaks, resection of telomere ends is not coordinated. Telomere restitution at permissive temperature is rapid (< 1 h) in ylcu70 Delta cdc13-1 cells. Surprisingly, survival can be high although strain background dependent. Given large amount of resected telomeres, we examined associated proteins. Up to 90% of cells have >= 1 Rfal (RPA) focus and 60% have multiple foci when similar to 30-40 telomeres/cell are resected. The ends are dispersed in the nucleus suggesting wide distribution of resected telomeres across nuclear space. The previously reported Rad52 nuclear centers of repair for random DSBs also appear in cells with many resected telomere ends, suggesting a Rad52 commonality to the organization of single strand ends and/or limitation on interactions of single-strand ends with Rad52.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.3
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据