SCFDia2 regulates DNA replication forks during S-phase in budding yeast
SCFDia2 regulates DNA replication forks during S-phase in budding yeast
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DOI：
10.1038/emboj.2009.320
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发表时间：
2009-12-02
期刊：
影响因子：
11.4
通讯作者：
Kamura, Takumi
中科院分区：
文献类型：
--
作者：
Mimura, Satoru;Komata, Makiko;Kamura, Takumi
Dia 2是一种F-box蛋白，其参与芽殖酵母酿酒酵母（Saccharomyces cerevisiae）中DNA复制的调节。然而，Dia 2的功能在很大程度上仍然未知。在这项研究中，我们报告说，Dia 2与复制叉和调节复制叉进展。使用改进的酵母双杂交筛选，我们已经确定了复制体的组成部分（Mrc 1，Ctf 4和Mcm 2），作为Dia 2结合蛋白。Mrc 1和Ctf 4在体内和体外均被SCFDia 2泛素化。Dia 2的结构域分析表明，富含亮氨酸的重复基序是不可或缺的调节复制体的进展，而tetratricopeptide repeat（TPR）基序参与与复制体组件的相互作用。此外，TPR基序被证明参与Dia 2稳定性;删除TPR稳定Dia 2，模拟DNA损伤的效果。ChIP芯片分析表明，Dia 2定位于复制叉，并调节叉的进展对羟基脲治疗。这些结果表明，Dia 2通过与复制体组分的直接相互作用参与复制体活性的调节。The EMBO Journal（2009）28，3693-3705. doi：10.1038/doj.2009.320; 2009年11月12日在线发布
Dia2 is an F-box protein, which is involved in the regulation of DNA replication in the budding yeast Saccharomyces cerevisiae. The function of Dia2, however, remains largely unknown. In this study, we report that Dia2 is associated with the replication fork and regulates replication fork progression. Using modified yeast two-hybrid screening, we have identified components of the replisome (Mrc1, Ctf4 and Mcm2), as Dia2-binding proteins. Mrc1 and Ctf4 were ubiquitinated by SCFDia2 both in vivo and in vitro. Domain analysis of Dia2 revealed that the leucine-rich repeat motif was indispensable for the regulation of replisome progression, whereas the tetratricopeptide repeat (TPR) motif was involved in the interaction with replisome components. In addition, the TPR motif was shown to be involved in Dia2 stability; deleting the TPR stabilized Dia2, mimicking the effect of DNA damage. ChIP-on-chip analysis illustrated that Dia2 localizes to the replication fork and regulates fork progression on hydroxyurea treatment. These results demonstrate that Dia2 is involved in the regulation of replisome activity through a direct interaction with replisome components. The EMBO Journal (2009) 28, 3693-3705. doi: 10.1038/emboj.2009.320; Published online 12 November 2009