Different requirements for the association of ATR-ATRIP and 9-1-1 to the stalled replication forks
Different requirements for the association of ATR-ATRIP and 9-1-1 to the stalled replication forks
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DOI：
10.1016/j.gene.2006.03.019
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发表时间：
2006-08-01
期刊：
影响因子：
3.5
通讯作者：
Shirahige, Katsuhiko
中科院分区：
文献类型：
--
作者：
Kanoh, Yutaka;Tamai, Katsuyuki;Shirahige, Katsuhiko
DNA复制检查点作为S期进程的一种监控机制，对维持基因组的完整性起着至关重要的作用。多种因子已被表征为参与检查点信号转导。RPA是一种单链DNA结合蛋白，被发现负责形成被检查点传感器识别的结构，然后发出激活DNA损伤检查点的初始信号。在这里，我们使用的rpal基因的突变体，rfal-t11，有缺陷的招募检查点传感器蛋白的双链断裂的网站，检查突变体的DNA复制检查点的激活的影响。我们发现，突变体细胞激活DNA复制检查点正常，并显示没有缺陷，招募ATR-ATRIP，一个主要的传感器复合物，是必不可少的DNA复制/损伤检查点，停止叉的网站。相比之下，突变体在将9-1-1复合物（另一种在DNA损伤检查点信号转导中起作用的传感器复合物）募集到停滞的叉上方面有缺陷。此外，我们还发现缺失Mrc 1后，rfal-t11突变体对Ffu的敏感性明显增加，而缺失损伤检查点特异性衔接子Rad 9对Ffu的敏感性影响不大。这些数据有力地表明，rfal-t11突变体主要是在激活DNA损伤检查点方面有缺陷，并且ATR-ATRIP和9-1-1向应激叉募集的分子需求可能不同。(c)2006 Elsevier B. V.保留所有权利。
DNA replication checkpoint, a surveillance mechanism for S-phase progression, plays a crucial role for the maintenance of genome integrity. A variety of factors have been characterized to be involved in the checkpoint signal transduction. Rpa, a single strand DNA binding protein, was found to be responsible for forming a structure that is recognized by checkpoint sensors and then emits the initial signal for the activation of DNA damage checkpoint. Here we use a mutant of rpal gene, rfal-t11, that has defects in recruiting checkpoint sensor proteins to the site of double strand break, to examine the mutant's effects on the activation of DNA replication checkpoint. We found that the mutant cells activated DNA replication checkpoint normally and showed no defects in recruiting ATR-ATRIP, a major sensor complex that is essential for DNA replication/damage checkpoint, to the site of stalled forks. In contrast, the mutant was defective in recruiting 9-1-1 complex, another sensor complex that functions in DNA damage checkpoint signal transduction, to the stalled forks. Moreover we found that sensitivity for Ffu obviously appeared in rfal-t11 mutant when Mrc1 was deleted, while deletion of Rad9, an adaptor specific for damage checkpoint, had subtle effect. These data strongly suggest that rfal-t11 mutant was mainly defective for activating DNA damage checkpoint and molecular requirement for the recruitment of ATR-ATRIP and 9-1-1 to the stressed forks may be different. (c) 2006 Elsevier B.V. All rights reserved.