Biosynthesis and properties of the adenovirus 2 L1-encoded 52,000- and 55,000-Mr proteins.
Biosynthesis and properties of the adenovirus 2 L1-encoded 52,000- and 55,000-Mr proteins.
复制标题
腺病毒 2 L1 编码的 52,000- 和 55,000-Mr 蛋白的生物合成和特性。
DOI：
10.1128/jvi.57.3.839-847.1986
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发表时间：
1986
影响因子：
5.4
通讯作者：
Green,M
中科院分区：
文献类型：
--
作者：
Lucher,LA;Symington,JS;Green,M
2型腺病毒L1区位于病毒基因组上30.7~39.2个MAP单位，在病毒复制的早期和晚期由主要的晚期启动子转录而来，在体外L1特异的RNA上翻译了一对52,000-MR(52K)蛋白-55K蛋白。为了研究L152K和55K蛋白的生物合成和性质，我们制备了针对预测的N末端附近的合成肽的抗体。免疫沉淀和免疫印迹分析表明，该抗体识别2型腺病毒感染细胞合成的主要52K和55K蛋白，与体外翻译的52K-55K蛋白相似。肽图分析表明，免疫沉淀的52K和55K蛋白具有非常密切的亲缘关系。这两个L1蛋白都被磷酸化了，而且它们在相似的位置被磷酸化。脉冲追逐分析没有发现52K和55K蛋白之间的前体-产物关系。L152K和55K蛋白的生物合成在感染后6~7h开始，在早期的1A区和1B19K(175R)T抗原的生物合成之后，在15h左右达到最大速率，最大速率保持到感染后至少25h。在任何时候，55K蛋白的合成水平似乎都比52K蛋白高出几倍。这两种蛋白都相当稳定，并一直积累到感染后的后期。病毒DNA复制对L1蛋白的形成不是必需的。因此，L1 52K-55K基因似乎以一种不同于经典的早期和晚期病毒基因的方式受到调控，但与I-Leader(Symington等人，J.Virol)编码的蛋白质相似。57：849-856,1986)。免疫荧光显微镜检测到L1蛋白位于细胞核内，免疫印迹分析发现L1蛋白主要与核膜结合。
The adenovirus type 2 L1 region, which is located at 30.7 to 39.2 map units on the viral genome, is transcribed from the major late promoter during both early and late stages of virus replication, and a 52,000-Mr (52K) protein-55K protein doublet has been translated in vitro on L1-specific RNA. To investigate the biosynthesis and properties of the L1 52K and 55K proteins, we prepared antibody against a synthetic peptide encoded near the predicted N terminus. As determined by immunoprecipitation and immunoblot analysis, the antipeptide antibody recognized major 52K and 55K proteins synthesized in adenovirus type 2-infected cells that appeared to be identical to the 52K-55K doublet translated in vitro. The immunoprecipitated 52K and 55K proteins were very closely related, as shown by a peptide map analysis. Both L1 proteins were phosphorylated, and they were phosphorylated at similar sites. No precursor-product relationship was detected between the 52K and 55K proteins by a pulse-chase analysis. Biosynthesis of the L1 52K and 55K proteins began about 6 to 7 h postinfection, after biosynthesis of the early region 1A and early region 1B 19K (175R) T antigens, and reached a maximum rate at about 15 h; the maximum rate was maintained until at least 25 h postinfection. At all times, the 55K protein appeared to be synthesized at a severalfold-higher level than the 52K protein. Both proteins were quite stable and accumulated until late times after infection. Viral DNA replication was not essential for formation of the L1 proteins. Thus, the L1 52K-55K gene appears to be regulated in a manner different from the classical early and late viral genes but similar to the protein encoded by the i-leader (Symington et al., J. Virol. 57:849-856, 1986). The L1 proteins were detected in the cell nucleus by immunofluorescence microscopy with antipeptide antibody and were found to be primarily associated with the nuclear membrane by an immunoblot analysis of subcellular fractions.