Partin K, Zybarth G, Ehrlich L, DeCrombrugghe M, Wimmer E, Carter C
Department of Microbiology, State University of New York, Stony Brook 11794.
Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4776-80. doi: 10.1073/pnas.88.11.4776.
Human immunodeficiency virus type 1 expresses structural proteins and replicative enzymes within gag and gag-pol precursor polyproteins. Specific proteolytic processing of the precursors by the viral proteinase is essential for maturation of infectious viral particles. We have studied the activity of proteinase in its immature form, as part of a gag-pol fusion protein, in an in vitro expression system. We found that deletion of p6*, the region in pol upstream of proteinase, resulted in improved processing of the precursor. A modified proteinase is released, but it functions less efficiently than wild type. Improved autoprocessing correlates with increased accessibility of the active site region in the polyprotein carrying the p6* deletion. Our results suggest that p6* is involved in the regulation of proteinase activation, perhaps as a region limiting the interaction of the active site and substrate binding domain with the remainder of the polyprotein. Release of p6* inhibition may be an activation step necessary for infectious particle maturation.
1型人类免疫缺陷病毒在gag和gag-pol前体多聚蛋白中表达结构蛋白和复制酶。病毒蛋白酶对前体进行特定的蛋白水解加工对于感染性病毒颗粒的成熟至关重要。作为gag-pol融合蛋白的一部分,我们在体外表达系统中研究了未成熟形式的蛋白酶活性。我们发现,缺失蛋白酶上游pol区的p6*,可改善前体的加工。释放出一种修饰的蛋白酶,但其功能效率低于野生型。自加工的改善与携带p6缺失的多聚蛋白中活性位点区域的可及性增加相关。我们的结果表明,p6参与蛋白酶激活的调节,可能作为一个限制活性位点和底物结合结构域与多聚蛋白其余部分相互作用的区域。释放p6*抑制可能是感染性颗粒成熟所必需的激活步骤。