Kingsman S M, Smith M D, Samuel C E
Proc Natl Acad Sci U S A. 1980 May;77(5):2419-23. doi: 10.1073/pnas.77.5.2419.
The effect of interferon treatment on proteins synthesized in simian virus 40 (SV40)-infected cells in the presence of cytosine arabinoside was investigated. The following results were obtained: (i) In addition to previously described large tumor (T) antigen (94 kilodaltons) and small tumor (t) antigen (19 kilodaltons), a 62-kilodalton polypeptide was immunoprecipitated by SV40 anti-T antiserum from extracts of infected CV-1 and BSC-1 monkey kidney cells and transformed SV3T3 mouse cells. The 94-, 62-, and 19-kilodalton polypeptides were not precipitated with normal serum from extracts of infected cells, and they were not present in extracts of uninfected cells. (ii) The de novo synthesis of the 94-, 62-, and 19-kilodalton tumor antigens was inhibited in CV-1 and BSC-1 cells treated with interferon before infection; total cellular protein synthesis was not significantly affected by interferon treatment. The relative interferon sensitivity of the three polypeptides in lytically infected monkey cells was comparable; by contrast, interferon did not affect their synthesis in transformed mouse cells. (iii) The 62-kilodalton polypeptide was detected in monkey cells infected with the following strains of SV40: tsA30 at both 33 degrees C and 41 degrees C; wt 708, the parent of tsA30; dI 884; and wt 830, the parent of dI 884. The amount of the 62-kilodalton species relative to T antigen was significantly greater in tsA30-infected cells as compared to cells infected with other SV40 strains. (iv) T, t, and 62-kilodalton polypeptides were readily labeled with [(35)S]methionine during a 10-min pulse; in a subsequent chase, the (35)S-labeled 94-kilodalton T antigen was apparently converted to 89- and 84-kilodalton polypeptides but not to either the 62-kilodalton polypeptide species or t antigen. (v) Partial peptide maps suggest that the 62-kilodalton polypeptide and T antigen are closely related. (vi) In addition to the above described 62-kilodalton polypeptide, a 54-kilodalton polypeptide was also detected. However, the 54-kilodalton species appears to be of cellular origin because it was immunoprecipitated with both normal and anti-T antiserum from uninfected and lytically infected cells and from virally transformed cells.
研究了在阿糖胞苷存在的情况下,干扰素处理对猿猴病毒40(SV40)感染细胞中合成的蛋白质的影响。得到了以下结果:(i)除了先前描述的大肿瘤(T)抗原(94千道尔顿)和小肿瘤(t)抗原(19千道尔顿)外,用SV40抗T抗血清从感染的CV-1和BSC-1猴肾细胞以及转化的SV3T3小鼠细胞提取物中免疫沉淀出一种62千道尔顿的多肽。感染细胞提取物中的正常血清不能沉淀94、62和19千道尔顿的多肽,未感染细胞的提取物中也不存在这些多肽。(ii)在感染前用干扰素处理的CV-1和BSC-1细胞中,94、62和19千道尔顿肿瘤抗原的从头合成受到抑制;干扰素处理对总细胞蛋白质合成没有显著影响。在裂解感染的猴细胞中,这三种多肽对干扰素的相对敏感性相当;相比之下,干扰素不影响它们在转化小鼠细胞中的合成。(iii)在感染以下SV40毒株的猴细胞中检测到了62千道尔顿的多肽:tsA30在33℃和41℃时均有;wt 708,tsA30的亲本;dI 884;以及dI 884的亲本wt 830。与感染其他SV40毒株的细胞相比,tsA30感染细胞中62千道尔顿物种相对于T抗原的量显著更多。(iv)在10分钟的脉冲期间,T、t和62千道尔顿的多肽很容易被[³⁵S]甲硫氨酸标记;在随后的追踪中,³⁵S标记的94千道尔顿T抗原显然转化为89和84千道尔顿的多肽,但没有转化为62千道尔顿的多肽物种或t抗原。(v)部分肽图表明62千道尔顿的多肽与T抗原密切相关。(vi)除了上述62千道尔顿的多肽外,还检测到一种54千道尔顿的多肽。然而,54千道尔顿的物种似乎起源于细胞,因为它在用正常血清和抗T抗血清从未感染和裂解感染的细胞以及病毒转化细胞中进行免疫沉淀时均被检测到。