Ide T, Whelly S, Baserga R
Proc Natl Acad Sci U S A. 1977 Aug;74(8):3189-92. doi: 10.1073/pnas.74.8.3189.
T-Antigen was partially purified from nuclei of cells transformed by simian virus 40 (SV 40). When nuclei isolated from either rat liver or quiescent hamster cells were preincubated with T-antigen preparations, there was a marked stimulation of RNA synthesis in an in vitro assay, up to 150% above control levels. The stimulation of RNA synthesis was inhibited by hamster antiserum against T-antigen but not by normal hamster serum. When the T-antigen preparations were fractionated on glycerol gradients, the fractions containing complement-fixing activity with antiserum to T-antigen also had the highest stimulatory activity on nuclear RNA synthesis. T-Antigen was also partially purified from nuclei of cells transformed by a temperature-sensitive A mutant of SV40. When preincubated up to 2 hr at 50 degrees, the T-antigen preparation from these temperature-sensitive A mutants was rapidly inactivated, in terms of both complement-fixing activity and ability to stimulate RNA synthesis in isolated rat liver nuclei. Under the same conditions of preincubation, T-antigen preparations from cells transformed by wild-type SV40 maintained their complement-fixing activity and ability to stimulate RNA synthesis. These results suggest that the biological action of T-antigen may be exerted at the level of transcription.
T抗原是从被猴病毒40(SV40)转化的细胞的细胞核中部分纯化得到的。当从大鼠肝脏或静止的仓鼠细胞中分离出的细胞核与T抗原制剂进行预孵育时,在体外测定中RNA合成有显著刺激,比对照水平高出150%。RNA合成的刺激被仓鼠抗T抗原血清抑制,但不被正常仓鼠血清抑制。当T抗原制剂在甘油梯度上进行分级分离时,含有与抗T抗原血清的补体结合活性的级分对核RNA合成也具有最高的刺激活性。T抗原也从被SV40温度敏感A突变体转化的细胞的细胞核中部分纯化得到。当在50℃预孵育长达2小时时,来自这些温度敏感A突变体的T抗原制剂在补体结合活性和刺激分离的大鼠肝脏细胞核中RNA合成的能力方面迅速失活。在相同的预孵育条件下,来自被野生型SV40转化的细胞的T抗原制剂保持其补体结合活性和刺激RNA合成的能力。这些结果表明T抗原的生物学作用可能在转录水平发挥。