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猿猴病毒40肿瘤特异性蛋白:在感染无缺陷腺病毒2型-猿猴病毒40杂交病毒的HeLa细胞中的亚细胞分布和代谢稳定性

Simian virus 40 tumor-specific proteins: subcellular distribution and metabolic stability in HeLa cells infected with nondefective adenovirus type 2-simian virus 40 hybrid viruses.

作者信息

Deppert W, Walter G, Linke H

出版信息

J Virol. 1977 Mar;21(3):1170-86. doi: 10.1128/JVI.21.3.1170-1186.1977.

Abstract

HeLa cells infected with adenovirus type 2 (Ad2)-simian virus 40 (SV40) hybrid viruses produce several SV40-specific proteins. These include the previously reported 28,000-dalton protein of Ad2+ND1, and 42,000- and 56,000-dalton proteins of Ad2+ND2, the 56,000-dalton protein of Ad2+ND4, and the 42,000-dalton protein of Ad2+ND5. In this report, we extend the list of SV40-specific proteins induced by Ad2+ND4 to include proteins of apparent molecular weights of 28,000 42,000, 60,000, 64,000, 72,000, 74,000, and a doublet of 95,000. Cell fractionation studies demonstrate that the SV40-specific proteins are detectable in the nuclear, cytoplasmic, and plasma membrane fractions. By pulse-chase and cell fractionation experiments, three classes of SV40-specific proteins can be distinguished with regard to metabolic stability: (i) unstable in the cytoplasmic but stable in the nuclear and plasma membrane fractions; (ii) stable in the nuclear, cytoplasmic, and plasma membrane fractions; and (iii) unstable in all subcellular fractions. Immunoprecipitation of infected cell extracts demonstrates that most of the above proteins share antigenic determinants with proteins expressed in hamsters bearing SV40-induced tumors. Only the 42,000-dalton protein of Ad2+ND5 is not immunoprecipitable.

摘要

感染2型腺病毒(Ad2)-猴病毒40(SV40)杂交病毒的HeLa细胞会产生几种SV40特异性蛋白。这些蛋白包括先前报道的Ad2 + ND1的28,000道尔顿蛋白、Ad2 + ND2的42,000道尔顿和56,000道尔顿蛋白、Ad2 + ND4的56,000道尔顿蛋白以及Ad2 + ND5的42,000道尔顿蛋白。在本报告中,我们将Ad2 + ND4诱导产生的SV40特异性蛋白列表进行了扩展,使其包括表观分子量为28,000、42,000、60,000、64,000、72,000、74,000的蛋白以及一个95,000的双峰。细胞分级分离研究表明,在细胞核、细胞质和质膜分级分离物中可检测到SV40特异性蛋白。通过脉冲追踪和细胞分级分离实验,就代谢稳定性而言,可以区分出三类SV40特异性蛋白:(i)在细胞质中不稳定,但在细胞核和质膜分级分离物中稳定;(ii)在细胞核、细胞质和质膜分级分离物中稳定;(iii)在所有亚细胞分级分离物中都不稳定。感染细胞提取物的免疫沉淀表明,上述大多数蛋白与携带SV40诱导肿瘤的仓鼠中表达的蛋白具有共同的抗原决定簇。只有Ad2 + ND5的42,000道尔顿蛋白不能被免疫沉淀。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f339/515658/ecf6ee0d692f/jvirol00207-0354-a.jpg

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