Construction and analysis of simian virus 40 origins defective in tumor antigen binding and DNA replication.
Construction and analysis of simian virus 40 origins defective in tumor antigen binding and DNA replication.
复制标题
肿瘤抗原结合和DNA复制缺陷的猿猴病毒40起源的构建和分析。
DOI：
10.1073/pnas.77.11.6491
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发表时间：
1980
影响因子：
11.1
通讯作者：
Tjian,R
中科院分区：
文献类型：
--
作者：
Myers,RM;Tjian,R
我们将含有猴病毒40（SV40）DNA复制起点、早期启动子和肿瘤（T）抗原结合位点的311个碱基对的DNA片段插入细菌质粒中并进行克隆，该重组质粒pSV01在体外与纯化的T抗原结合，并在提供大T抗原时在猴细胞中复制。通过体外诱变，在pSV 01 DNA的起始序列中产生了一系列缺失突变。将这些突变DNA在猴细胞中的复制与它们与纯化的D2蛋白结合的能力进行比较。与D2蛋白结合缺陷的突变DNA在猴细胞中也表现出复制水平降低。这些发现提供了生物化学证据，表明SV40 DNA合成的起始可能涉及T抗原与复制起点序列的直接相互作用。
We have inserted a 311-base pair DNA fragment containing the simian virus 40 (SV40) origin of DNA replication, the early promoter, and the tumor (T) antigen binding sites into a bacterial plasmid and cloned it. This recombinant plasmid, pSV01, binds to a purified T antigen in vitro and replicates in monkey cells when supplied with large T antigen. A series of deletion mutations was generated in the origin sequences of pSV01 DNA by mutagenesis in vitro. The replication of these mutant DNAs in monkey cells was compared with their ability to bind to purified D2 protein. Mutant DNAs deficient in binding to D2 protein also exhibit reduced levels of replication in monkey cells. These findings provide biochemical evidence that the initiation of SV40 DNA synthesis may involve a direct interaction of T antigen with sequences at the origin of replication.