Protease cleavage of reovirus capsid protein mu 1/mu 1C is blocked by alkyl sulfate detergents, yielding a new type of infectious subvirion particle

被引:45
作者
Chandran, K
Nibert, ML
机构
[1] UNIV WISCONSIN, COLL AGR & LIFE SCI, DEPT BIOCHEM, MADISON, WI 53706 USA
[2] UNIV WISCONSIN, GRAD SCH, INST MOL VIROL, MADISON, WI 53706 USA
关键词
D O I
10.1128/JVI.72.1.467-475.1998
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Mammalian reovirus virions undergo partial disassembly of the outer capsid upon exposure to proteases in vitro, producing infectious subvirion particles (ISVPs) that lack protein sigma 3 and contain protein mu 1/mu 1C as endoprotease-generated fragments mu 1 delta/delta and phi. ISVPs are thought to be required for two early steps in reovirus infection: membrane penetration and activation of the particle-bound viral transcriptase complexes. Genetic and biochemical evidence implicates outer-capsid protein yl in both these steps. To determine whether the cleavage of mu 1/mu 1C is relevant to the unique properties of ISVPs, we analyzed the properties of novel subvirion particles that lacked sigma 3 yet retained mu 1/mu 1C in an uncleaved but cleavable form. These detergent-plus-protease subvirion particles (dpSVPs) were produced by treating virions with chymotrypsin in the presence of micelle-forming concentrations of alkyl sulfate detergents. Infections with dpSVPs in murine L or canine MDCK cells provided evidence that the cleavage of mu 1/mu 1C during viral entry into these cells is dispensable for reovirus infection. Additionally, dpSVPs behaved like ISVPs in their capacity to permeabilize lipid bilayers and to undergo transcriptase activation in vitro, supporting the conclusion that cleavage of mu 1/mu 1C to mu 1 delta/delta and phi during viral entry is not required for either membrane penetration or transcriptase activation in cells. The capacity of alkyl sulfate detergents to inhibit the cleavage of mu 1/mu 1C in a reversible fashion suggests a specific association between virus particle and detergent micelles that may mimic virus particle-phospholipid membrane interactions during reovirus entry into cells.
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页码:467 / 475
页数:9
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