Gustin K E, Lutz P, Imperiale M J
Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor 48109-0620, USA.
J Virol. 1996 Sep;70(9):6463-7. doi: 10.1128/JVI.70.9.6463-6467.1996.
The adenovirus L1 52/55-kDa protein is expressed both in the early and late stages of infection, raising the possibility that it has multiple roles in the viral life cycle. To obtain possible insights into these roles, the yeast two-hybrid system was used to examine the interactions of the 52/55-kDa protein with viral and cellular factors. cDNA expression libraries from human 293 cells at both early and late stages of adenovirus type 5 infection were constructed and screened, with the 52/55-kDa protein being used as bait. Characterization of positive clones revealed that the adenovirus IVa2 gene product interacted specifically with the 52/55-kDa protein. In addition, the IVa2 protein was shown to interact with a bacterial glutathione S-transferase-52/55-kDa fusion protein in vitro, further supporting the finding with the yeast two-hybrid system. Finally, coimmunoprecipitation studies confirmed that the 52/55-kDa protein and IVa2 polypeptide interact specifically during the course of adenovirus infection. A potential role for the IVa2-52/55-kDa protein interaction in the regulation of transcription from the major late promoter and in viral assembly is discussed.
腺病毒L1 52/55 kDa蛋白在感染的早期和晚期均有表达，这增加了其在病毒生命周期中具有多种作用的可能性。为了深入了解这些作用，利用酵母双杂交系统检测52/55 kDa蛋白与病毒及细胞因子的相互作用。构建并筛选了来自人293细胞在腺病毒5型感染早期和晚期的cDNA表达文库，以52/55 kDa蛋白作为诱饵。对阳性克隆的鉴定表明，腺病毒IVa2基因产物与52/55 kDa蛋白特异性相互作用。此外，IVa2蛋白在体外还显示出与细菌谷胱甘肽S-转移酶-52/55 kDa融合蛋白相互作用，进一步支持了酵母双杂交系统的研究结果。最后，免疫共沉淀研究证实，在腺病毒感染过程中，52/55 kDa蛋白与IVa2多肽特异性相互作用。本文讨论了IVa2-52/55 kDa蛋白相互作用在主要晚期启动子转录调控和病毒组装中的潜在作用。
