Reconstitution of simian virus 40 DNA replication with purified proteins.
Reconstitution of simian virus 40 DNA replication with purified proteins.
复制标题
用纯化的蛋白质重建猿猴病毒 40 DNA 复制。
DOI：
10.1073/pnas.87.22.8692
复制
发表时间：
1990
影响因子：
11.1
通讯作者：
Kelly,TJ
中科院分区：
文献类型：
--
作者：
Weinberg,DH;Collins,KL;Simancek,P;Russo,A;Wold,MS;Virshup,DM;Kelly,TJ
用7种高度纯化的细胞蛋白和SV40大肿瘤(T)抗原重建了含有类人猿病毒40 （SV40）起源DNA复制的质粒DNA分子。DNA合成的起始完全依赖于T抗原、复制蛋白A和DNA聚合酶-引物酶复合物，并由蛋白磷酸酶2A的催化亚基刺激。新生链的有效延伸还需要增殖细胞核抗原、复制因子C、DNA拓扑异构酶I和DNA聚合酶δ。电子显微镜研究表明，DNA复制始于病毒起源，并通过含有两个朝相反方向移动的分叉的中间体进行。这些发现表明，重组的复制反应具有真实病毒DNA复制的许多特征。
Replication of plasmid DNA molecules containing the simian virus 40 (SV40) origin of DNA replication has been reconstituted with seven highly purified cellular proteins plus the SV40 large tumor (T) antigen. Initiation of DNA synthesis is absolutely dependent upon T antigen, replication protein A, and the DNA polymerase alpha-primase complex and is stimulated by the catalytic subunit of protein phosphatase 2A. Efficient elongation of nascent chains additionally requires proliferating cell nuclear antigen, replication factor C, DNA topoisomerase I, and DNA polymerase delta. Electron microscopic studies indicate that DNA replication begins at the viral origin and proceeds via intermediates containing two forks that move in opposite directions. These findings indicate that the reconstituted replication reaction has many of the characteristics expected of authentic viral DNA replication.