Department of Microbiology and Infectious Diseases, Faculty of Medicine, Université de Sherbrooke, Sherbrooke, Quebec, Canada.
EMBO J. 2010 Sep 1;29(17):3007-19. doi: 10.1038/emboj.2010.155. Epub 2010 Jul 13.
The assembly of a protective cap onto the telomeres of eukaryotic chromosomes suppresses genomic instability through inhibition of DNA repair activities that normally process accidental DNA breaks. We show here that the essential Cdc13-Stn1-Ten1 complex is entirely dispensable for telomere protection in non-dividing cells. However, Yku and Rap1 become crucially important for this function in these cells. After inactivation of Yku70 in G1-arrested cells, moderate but significant telomere degradation occurs. As the activity of cyclin-dependent kinases (CDK) promotes degradation, these results suggest that Yku stabilizes G1 telomeres by blocking the access of CDK1-independent nucleases to telomeres. The results indeed show that both Exo1 and the Mre11/Rad50/Xrs2 complex are required for telomeric resection after Yku loss in non-dividing cells. Unexpectedly, both asynchronously growing and quiescent G0 cells lacking Rap1 display readily detectable telomere degradation, suggesting an earlier unanticipated function for this protein in suppression of nuclease activities at telomeres. Together, our results show a high flexibility of the telomeric cap and suggest that distinct configurations may provide for efficient capping in dividing versus non-dividing cells.
在真核染色体的端粒上组装保护帽可通过抑制通常处理意外 DNA 断裂的 DNA 修复活性来抑制基因组不稳定性。我们在这里表明,在非分裂细胞中,必不可少的 Cdc13-Stn1-Ten1 复合物对于端粒保护完全是可有可无的。然而,Yku 和 Rap1 在这些细胞中对于该功能变得至关重要。在 G1 期阻滞的细胞中失活 Yku70 后,端粒会发生中度但明显的降解。由于细胞周期蛋白依赖性激酶 (CDK) 的活性促进降解,这些结果表明 Yku 通过阻止 CDK1 非依赖性核酶接近端粒来稳定 G1 端粒。结果确实表明,在非分裂细胞中端粒切除后,Exo1 和 Mre11/Rad50/Xrs2 复合物都需要。出乎意料的是,缺乏 Rap1 的非同步生长和静止 G0 细胞都显示出可检测到的端粒降解,这表明该蛋白在抑制端粒处的核酶活性方面具有更早的预期功能。总之,我们的结果表明端粒帽具有很高的灵活性,并表明在有丝分裂和非有丝分裂细胞中,不同的构象可能提供有效的端粒封闭。