Identification of proteins that may directly interact with human RPA
Identification of proteins that may directly interact with human RPA
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DOI：
10.1093/jb/mvq085
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发表时间：
2010-11-01
影响因子：
2.7
通讯作者：
Ishimi, Yukio
中科院分区：
文献类型：
--
作者：
Nakaya, Ryou;Takaya, Junichiro;Ishimi, Yukio
RPA由三个亚基（RPA 1、2和3）组成，在DNA交易中发挥着重要作用。在DNA复制叉处，RPA与单链DNA区域结合以稳定结构并组装其他复制蛋白。RPA和几种复制蛋白之间的相互作用已被报道，但分析并不全面。我们系统地进行了定性分析，以识别RPA相互作用伙伴，以了解复制叉处的蛋白质-蛋白质相互作用。我们在昆虫细胞中表达了人RPA的三个亚基，以及一种复制蛋白，该蛋白在正常条件下和/或复制应激条件下存在于叉处，以检查相互作用。在总共检测的30种蛋白质中，发现至少有14种蛋白质与RPA相互作用。RPA与DNA复制起始和延伸过程中的MCM 3 -7、MCM-BP和CDC 45蛋白相互作用。RPA与TIPIN、CLASPIN和RAD 17结合，它们参与DNA复制检查点功能。RPA还与细胞周期蛋白依赖性激酶和负调节DNA复制的Rb蛋白的氨基末端片段结合。这些结果表明，RPA与在复制叉进展的调节中发挥作用的那些特定蛋白质相互作用。
RPA, which consisted of three subunits (RPA1, 2 and 3), plays essential roles in DNA transactions. At the DNA replication forks, RPA binds to single-stranded DNA region to stabilize the structure and to assemble other replication proteins. Interactions between RPA and several replication proteins have been reported but the analysis is not comprehensive. We systematically performed the qualitative analysis to identify RPA interaction partners to understand the protein-protein interaction at the replication forks. We expressed in insect cells the three subunits of human RPA, together with one replication protein, which is present at the forks under normal conditions and/or under the replication stress conditions, to examine the interaction. Among 30 proteins examined in total, it was found that at least 14 proteins interacted with RPA. RPA interacted with MCM3-7, MCM-BP and CDC45 proteins among the proteins that play roles in the initiation and the elongation of the DNA replication. RPA bound with TIPIN, CLASPIN and RAD17, which are involved in the DNA replication checkpoint functions. RPA also bound with cyclin-dependent kinases and an amino-terminal fragment of Rb protein that negatively regulates DNA replication. These results suggest that RPA interacts with the specific proteins among those that play roles in the regulation of the replication fork progression.