The S-cerevisiae Rrm3p DNA helicase moves with the replication fork and affects replication of all yeast chromosomes
The S-cerevisiae Rrm3p DNA helicase moves with the replication fork and affects replication of all yeast chromosomes
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DOI：
10.1101/gad.1478906
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发表时间：
2006-11-15
影响因子：
10.5
通讯作者：
Zakian, Virginia A.
中科院分区：
文献类型：
--
作者：
Azvolinsky, Anna;Dunaway, Stephen;Zakian, Virginia A.
酿酒酵母DNA解旋酶Rrm3p是通过分散在整个基因组中的> 1000个离散位点进行正常叉进展所需要的。在这里，我们表明，复制的所有酵母染色体显着延迟rrm3细胞。延迟复制，甚至在一个地区，缺乏任何预测的Rrm3p依赖性网站。基于在二维凝胶中的复制中间体的模式，除了在已知的Rrm3p依赖性位点之外，在rrm3细胞中叉运动的速率似乎与野生型相似。这些数据表明，尽管Rrm3p在DNA复制中具有全局作用，但其活性仅或主要在特定的难以复制的位点上需要。染色质免疫沉淀的标准，Rrm3p与Rrm3p依赖和独立的网站，并通过两者移动与复制叉。此外，Rrm3p与Pol2p，DNA聚合酶的催化亚基，在体内相互作用。因此，Rrm3p很可能是复制叉装置的一个组件，而不是在复制叉在这些站点停止时被招募到其作用站点。
The Saccharomyces cerevisiae DNA helicase Rrm3p is needed for normal fork progression through > 1000 discrete sites scattered throughout the genome. Here we show that replication of all yeast chromosomes was markedly delayed in rrm3 cells. Delayed replication was seen even in a region that lacks any predicted Rrm3p-dependent sites. Based on the pattern of replication intermediates in two-dimensional gels, the rate of fork movement in rrm3 cells appeared similar to wild-type except at known Rrm3p-dependent sites. These data suggest that although Rrm3p has a global role in DNA replication, its activity is needed only or primarily at specific, difficult-to-replicate sites. By the criterion of chromatin immunoprecipitation, Rrm3p was associated with both Rrm3p-dependent and -independent sites, and moved with the replication fork through both. In addition, Rrm3p interacted with Pol2p, the catalytic subunit of DNA polymerase epsilon, in vivo. Thus, rather than being recruited to its sites of action when replication forks stall at these sites, Rrm3p is likely a component of the replication fork apparatus.