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腺病毒相关病毒诱导的KB细胞中多肽的特性分析

Characterization of adenovirus-associated virus-induced polypeptides in KB cells.

作者信息

Buller R M, Rose J A

出版信息

J Virol. 1978 Jan;25(1):331-8. doi: 10.1128/JVI.25.1.331-338.1978.

Abstract

Electrophoretic analysis of KB cells coinfected with adenovirus-associated virus (AAV) type 2, a defective parvovirus, and adenovirus type 5 (as helper) have revealed the synthesis in vivo of at least five AAV-specific polypeptides. The three largest polypeptides, with molecular weights of 90,700, 71,600, and 60,000 comigrated in polyacrylamide gels with the three AAV structural polypeptides. The remaining two polypeptides had molecular weights of 24,900 and 15,800. The concentrations of the AAV-induced polypeptides relative to one another remained approximately constant during the infectious cycle, and the structural components were present in proportions similar to those found in purified virions. As determined by pulse-chase experiments, all polypeptides were generated at the level of protein synthesis and not by posttranslational proteolytic processing. Although inhibitors of proteolytic enzymes failed to influence the pattern of AAV-induced polypeptides, and amino acid analog, L-canavanine, blocked the appearances of both the major structural polypeptide (60,000 daltons) and the larger nonstructural polypeptide (24,900 daltons). Taken in conjunction with pulse-chase data, this result supports a model whereby the major virion polypeptide is produced by proteolytic cleavage of the nascent polypeptide chain.

摘要

用2型腺相关病毒(AAV,一种缺陷型细小病毒)和5型腺病毒(作为辅助病毒)共同感染KB细胞后进行的电泳分析显示,在体内至少合成了五种AAV特异性多肽。三种最大的多肽,分子量分别为90,700、71,600和60,000,在聚丙烯酰胺凝胶中与三种AAV结构多肽迁移率相同。其余两种多肽的分子量分别为24,900和15,800。在感染周期中,AAV诱导的多肽彼此之间的浓度大致保持恒定,并且结构成分的比例与在纯化病毒粒子中发现的比例相似。通过脉冲追踪实验确定,所有多肽都是在蛋白质合成水平产生的,而不是通过翻译后蛋白水解加工产生的。虽然蛋白水解酶抑制剂未能影响AAV诱导多肽的模式,但氨基酸类似物L-刀豆氨酸阻止了主要结构多肽(60,000道尔顿)和较大的非结构多肽(24,900道尔顿)的出现。结合脉冲追踪数据,这一结果支持了一个模型,即主要病毒粒子多肽是由新生多肽链的蛋白水解切割产生的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f7d4/353931/de48781d2e02/jvirol00193-0347-a.jpg

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