Yeast Rev1 protein promotes complex formation of DNA polymerase ζ with Pol32 subunit of DNA polymerase δ
Yeast Rev1 protein promotes complex formation of DNA polymerase ζ with Pol32 subunit of DNA polymerase δ
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DOI：
10.1073/pnas.0902175106
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发表时间：
2009-06-16
影响因子：
11.1
通讯作者：
Prakash, Satya
中科院分区：
文献类型：
--
作者：
Acharya, Narottam;Johnson, Robert E.;Prakash, Satya
酵母DNA聚合酶（Pol）δ是DNA复制所必需的，由3个亚基组成，Pol 3，Pol 31和Pol 32。其中，催化亚基Pol 3和第二亚基Pol 31是必需的，而Pol 32亚基对于DNA复制不是必需的。虽然Pol 32是Pol delta的组成部分，但它也是Pol zeta跨病变合成（TLS）所必需的。为了开始破译Pol 32参与Polzeta介导的TLS的基础，在这里，我们研究Pol 32是否与Polzeta或其相关蛋白质发生物理相互作用，并为Pol 32与Rev 1的物理相互作用提供证据。Rev 1在Pol zeta介导的TLS中起着不可或缺的结构作用，它结合Pol zeta的Rev 3催化亚基。在这里，我们表明，虽然Pol 32不直接结合Pol zeta，Pol 32可以通过与Rev 1的相互作用结合Rev 1-Pol zeta复合物。我们发现，Pol 32结合对DNA合成没有刺激作用，无论是Rev 1在Rev 1-Pol 32复合物中还是Pol zeta在Pol zeta-Rev 1-Pol 32复合物中，无论增殖细胞核抗原是否已加载到DNA上。我们讨论了证据的生物学意义的Rev 1结合Pol 32的Polzeta功能在TLS和建议的结构性作用Rev 1在调节结合Polzeta与Pol 32在Poldelta停滞在病变部位。
Yeast DNA polymerase (Pol) delta, essential for DNA replication, is comprised of 3 subunits, Pol3, Pol31, and Pol32. Of these, the catalytic subunit Pol3 and the second subunit Pol31 are essential, whereas the Pol32 subunit is not essential for DNA replication. Although Pol32 is an integral component of Pol delta, it is also required for translesion synthesis (TLS) by Pol zeta. To begin to decipher the bases of Pol32 involvement in Pol zeta-mediated TLS, here we examine whether Pol32 physically interacts with Pol zeta or its associated proteins and provide evidence for the physical interaction of Pol32 with Rev1. Rev1 plays an indispensable structural role in Pol zeta-mediated TLS and it binds the Rev3 catalytic subunit of Pol zeta. Here, we show that although Pol32 does not directly bind Pol zeta, Pol32 can bind the Rev1-Pol zeta complex through its interaction with Rev1. We find that Pol32 binding has no stimulatory effect on DNA synthesis either by Rev1 in the Rev1-Pol32 complex or by Pol zeta in the Pol zeta-Rev1-Pol32 complex, irrespective of whether proliferating cell nuclear antigen has been loaded onto DNA or not. We discuss evidence for the biological significance of Rev1 binding to Pol32 for Pol zeta function in TLS and suggest a structural role for Rev1 in modulating the binding of Pol zeta with Pol32 in Pol delta stalled at a lesion site.