Lin J Y, Simmons D T
School of Life and Health Sciences, University of Delaware, Newark 19716.
J Virol. 1991 Dec;65(12):6447-53. doi: 10.1128/JVI.65.12.6447-6453.1991.
Simian virus 40 (SV40) T antigen binds to the tumor suppressor p53 protein, and this association may contribute to oncogenic transformation by the virus. We investigated the importance of this binding on transformation by examining three replication-competent mutants of SV40 (402DE, 402DN, and 402DH). These mutants express T antigens defective in binding to human and monkey p53s but retain some binding with mouse p53. All showed significant reduction in their ability to induce transformed cell foci of two normal human cell lines as well as a slight reduction with mouse embryo cells. Other comparable mutants which express T antigens retaining the ability to complex with p53 were able to induce foci at wild-type levels in both human and mouse cells. Further studies were performed with five T-antigen-positive clones isolated from the few human cell foci that appeared after transfection with 402 mutant DNAs. All five clones reached senescence at about the same point as did the parental untransformed cells. However, six other human cell clones obtained after transfection with DNA from nondefective mutants or wild-type virus were still growing well at more than 10 passages beyond their expected life span. These results suggest that the ability of T antigen to form stable complexes with p53 is necessary for SV40 to extend the life span and partially transform human cells in culture.
猴病毒40(SV40)T抗原与肿瘤抑制蛋白p53结合,这种结合可能有助于该病毒的致癌转化。我们通过检测SV40的三个复制能力突变体(402DE、402DN和402DH)来研究这种结合对转化的重要性。这些突变体表达的T抗原在与人及猴p53结合方面存在缺陷,但仍与小鼠p53有一定结合。所有这些突变体在诱导两种正常人细胞系形成转化细胞集落的能力上均显著降低,对小鼠胚胎细胞的诱导能力也略有降低。其他表达能与p53形成复合物的T抗原的可比突变体,在人和小鼠细胞中均能以野生型水平诱导形成集落。我们对从用402突变体DNA转染后出现的少数人细胞集落中分离得到的五个T抗原阳性克隆进行了进一步研究。所有五个克隆与未转化的亲代细胞在大约相同的时间点进入衰老状态。然而,用无缺陷突变体或野生型病毒的DNA转染后获得的另外六个人类细胞克隆,在超过其预期寿命的10代以上仍生长良好。这些结果表明,T抗原与p53形成稳定复合物的能力对于SV40延长培养的人类细胞寿命和部分转化人类细胞是必要的。