Grussenmeyer T, Carbone-Wiley A, Scheidtmann K H, Walter G
Institut für Immunbiologie, Universität Freiburg, Federal Republic of Germany.
J Virol. 1987 Dec;61(12):3902-9. doi: 10.1128/JVI.61.12.3902-3909.1987.
Affinity-purified medium T antigen of wild-type polyomavirus and dl8, a transforming mutant with a deletion in the medium T gene, is associated with three cellular proteins with apparent molecular weights of 88,000 (88K protein), 61,000 (61K protein), and 37,000 (37K protein). Medium T antigen encoded by the nontransforming hrt mutants fails to associate with these proteins, whereas medium T antigen of the nontransforming mutant dl1015 is able to do so. Medium T antigen of the nontransforming mutant dl23 binds to the 61K and 37K proteins; however, binding to the 88K protein is uncertain. The pattern of complex formation between these proteins and medium T antigen resembles that of pp60c-src and medium T antigen. The binding of medium T antigen to the 88K, 61K, and 37K proteins, as well as to pp60c-src, might represent a necessary but insufficient step in transformation. By mixing extracts from infected and uninfected cells, complex formation between medium T antigen and the 88K, 61K, and 37K proteins can be demonstrated in vitro. Pulse-chase experiments indicated that in vivo the association between medium T antigen and the 61K and 37K proteins is a slow process. The latter two proteins are probably bound to each other in uninfected cells. On two-dimensional gels of whole-cell extract, the 61K protein comigrated with a minor protein with an isoelectric point of 5.2. The 61K protein was neither phosphorylated nor glycosylated. Polyomavirus tumor serum precipitated the 61K and 37K proteins independently of medium T antigen. Therefore, the 61K protein or the 37K protein or both have the properties of a cellular tumor antigen.
野生型多瘤病毒的亲和纯化中T抗原以及dl8(一种中T基因缺失的转化突变体)与三种细胞蛋白相关,这些蛋白的表观分子量分别为88,000(88K蛋白)、61,000(61K蛋白)和37,000(37K蛋白)。非转化hrt突变体编码的中T抗原无法与这些蛋白结合,而未转化突变体dl1015的中T抗原却能够结合。未转化突变体dl23的中T抗原能与61K和37K蛋白结合;然而,与88K蛋白的结合情况尚不确定。这些蛋白与中T抗原之间形成复合物的模式类似于pp60c-src与中T抗原之间的模式。中T抗原与88K、61K和37K蛋白以及pp60c-src的结合,可能是转化过程中一个必要但不充分的步骤。通过混合感染细胞和未感染细胞的提取物,可在体外证明中T抗原与88K、61K和37K蛋白之间形成复合物。脉冲追踪实验表明,在体内中T抗原与61K和37K蛋白之间的结合是一个缓慢的过程。后两种蛋白在未感染细胞中可能彼此结合。在全细胞提取物的二维凝胶上,61K蛋白与一种等电点为5.2的次要蛋白迁移位置相同。61K蛋白既不被磷酸化也不被糖基化。多瘤病毒肿瘤血清能独立于中T抗原沉淀61K和37K蛋白。因此,61K蛋白或37K蛋白或两者都具有细胞肿瘤抗原的特性。