THE CELLULAR DNA-POLYMERASE ALPHA-PRIMASE IS REQUIRED FOR PAPILLOMAVIRUS DNA-REPLICATION AND ASSOCIATES WITH THE VIRAL E1 HELICASE
THE CELLULAR DNA-POLYMERASE ALPHA-PRIMASE IS REQUIRED FOR PAPILLOMAVIRUS DNA-REPLICATION AND ASSOCIATES WITH THE VIRAL E1 HELICASE
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DOI：
10.1073/pnas.91.18.8700
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发表时间：
1994-08-30
影响因子：
11.1
通讯作者：
MOHR, IJ
中科院分区：
文献类型：
--
作者：
PARK, P;COPELAND, W;MOHR, IJ
乳头瘤病毒的持续感染涉及病毒DNA作为核质粒的维持，其复制需要宿主DNA聚合酶。细胞DNA聚合酶α-引发酶全酶的作用进行了探测，通过使用从啮齿动物细胞，复制牛乳头瘤病毒1和人乳头瘤病毒6 b DNA的病毒E1解旋酶和E2转录因子的存在下，可溶性提取物。针对聚合酶α的催化性180-kDa亚基的单克隆抗体抑制该系统中的DNA合成。向中和的提取物中加入纯化的人聚合酶α-引发酶全酶可恢复其DNA合成活性。E1的氨基末端424个氨基酸与p180聚合酶亚基形成特异性蛋白复合物。免疫复合物可以用含有DNA聚合酶活性的针对E1的抗体分离。此外，这种聚合酶活性可以被抗聚合酶α抗体中和。在这个体外系统中没有遇到perceptide屏障，因为牛E1可以与鼠和人的复制装置相互作用。尽管猿猴病毒40和多瘤病毒编码的大型肿瘤抗原与乳头瘤病毒E1蛋白具有有限的一级序列同源性，但功能蛾在一级蛋白质结构水平上的组织非常相似。除了它们的起源特异性DNA结合活性，这些解旋酶中的每一种都可以发挥作用，以帮助招募细胞聚合酶α-引发酶复合物到病毒复制起点。
Persistent infection by papillomaviruses involves the maintenance of viral DNA as a nuclear plasmid, the replication of which requires host DNA polymerases. The role of the cellular DNA polymerase alpha-primase holoenzyme was probed by using soluble extracts from rodent cells that replicate bovine papilloma virus 1 and human papilloma virus 6b DNA in the presence of the viral E1 helicase and the E2 transcription factor. Monoclonal antibodies directed against the catalytic 180-kDa subunit of polymerase alpha inhibit DNA synthesis in this system. Addition of purified human polymerase alpha-primase holoenzyme to neutralized extracts restores their DNA synthetic activity. The amino-terminal 424 amino acids of E1 forms a specific protein complex with the p180 polymerase subunit. Immune complexes can be isolated with antibodies directed against E1 that contain a DNA polymerase activity. Moreover, this polymerase activity tan be neutralized by anti-polymerase alpha antibodies. Permissivity barriers were not encountered in this in vitro system, as bovine E1 can interface with the murine and human replication apparatus. Although the large tumor antigens encoded by simian virus 40 and polyoma share limited primary sequence homology with the papillomavirus E1 proteins, the organization of functional moths at the level of primary protein structure is remarkably similar. In addition to their origin-specific DNA-binding activity, each of these helicases may function to help recruit the cellular polymerase alpha-primase complex to the-viral replication origin.