Interaction of hREV1 with three human Y-family DNA polymerases
Interaction of hREV1 with three human Y-family DNA polymerases
复制标题
DOI：
10.1111/j.1356-9597.2004.00747.x
复制
发表时间：
2004-06-01
期刊：
影响因子：
2.1
通讯作者：
Ohmori, H
中科院分区：
文献类型：
--
作者：
Ohashi, E;Murakumo, Y;Ohmori, H
Polkappa是参与跨损伤DNA合成（TLS）的许多DNA聚合酶之一。它与Poleta、Poliota和hREV 1一起属于聚合酶的Y家族沿着。与着色性干皮病变体（XPV）基因编码的Poleta不同，Polkappa不能绕过体外UV诱导的DNA损伤，但它能够准确有效地绕过苯并[a]芘（B[a]P）加合的鸟嘌呤。在试图鉴定将Polkappa靶向其同源DNA损伤的因子中，我们通过使用酵母双杂交测定来搜索Polkappa相互作用蛋白。我们发现Polkappa与hREV 1的C-末端区域相互作用。Poleta和Poliota也被发现与hREV 1的相同区域相互作用。通过pull-down和免疫共沉淀试验证实Polkappa和hREV 1之间的相互作用。已知hREV 1的C-末端区域与hREV 7相互作用，hREV 7是Polzeta的非催化亚基，是另一种结构上不相关的TLS酶，我们表明Polkappa和hREV 7与hR.EV1的相同C-末端区域结合。因此，我们的研究结果表明，hREV 1在跨损伤DNA合成的多酶，多步骤过程中起着关键作用。
Polkappa is one of many DNA polymerases involved in translesion DNA synthesis (TLS). It belongs to the Y-family of polymerases along with Poleta, Poliota and hREV1. Unlike Poleta encoded by the xeroderma pigmentosum variant (XPV) gene, Polkappa is unable to bypass UV-induced DNA damage in vitro, but it is able to bypass benzo[a]pyrene (B[a]P)-adducted guanines accurately and efficiently. In an attempt to identify factor(s) targeting Polkappa to its cognate DNA lesion(s), we searched for Polkappa-interacting proteins by using the yeast two-hybrid assay. We found that Polkappa interacts with a C-terminal region of hREV1. Poleta and Poliota were also found to interact with the same region of hREV1. The interaction between Polkappa and hREV1 was confirmed by pull-down and co-immunoprecipitation assays. The C-terminal region of hREV1 is known to interact with hREV7, a non-catalytic subunit of Polzeta that is another structurally unrelated TLS enzyme, and we show that Polkappa and hREV7 bind to the same C-terminal region of hR.EV1. Thus, our results suggest that hREV1 plays a pivotal role in the multi-enzyme, multi-step process of translesion DNA synthesis.