Reovirus replication protein μ2 influences cell tropism by promoting particle assembly within viral inclusions.

Ooms LS, Jerome WG, Dermody TS, Chappell JD
J Virol. 2012 86 (20): 10979-87

PMID: 22837214 · PMCID: PMC3457141 · DOI:10.1128/JVI.01172-12

The double-stranded RNA virus mammalian reovirus displays broad cell, tissue, and host tropism. A critical checkpoint in the reovirus replication cycle resides within viral cytoplasmic inclusions, which are biosynthetic centers of genome multiplication and new-particle assembly. Replication of strain type 3 Dearing (T3) is arrested in Madin-Darby canine kidney (MDCK) cells at a step subsequent to inclusion development and prior to formation of genomic double-stranded RNA. This phenotype is primarily regulated by viral replication protein μ2. To understand how reovirus inclusions differ in productively and abortively infected MDCK cells, we used confocal immunofluorescence and thin-section transmission electron microscopy (TEM) to probe inclusion organization and particle morphogenesis. Although no abnormalities in inclusion morphology or viral protein localization were observed in T3-infected MDCK cells using confocal microscopy, TEM revealed markedly diminished production of mature progeny virions. T3 inclusions were less frequent and smaller than those formed by T3-T1M1, a productively replicating reovirus strain, and contained decreased numbers of complete particles. T3 replication was enhanced when cells were cultivated at 31°C, and inclusion ultrastructure at low-temperature infection more closely resembled that of a productive infection. These results indicate that particle assembly in T3-infected MDCK cells is defective, possibly due to a temperature-sensitive structural or functional property of μ2. Thus, reovirus cell tropism can be governed by interactions between viral replication proteins and the unique cell environment that modulate efficiency of particle assembly.

MeSH Terms (17)

Animals Cell Line Dogs Inclusion Bodies, Viral Madin Darby Canine Kidney Cells Mice Microscopy, Electron, Transmission Phenotype Reoviridae RNA, Double-Stranded RNA, Viral Temperature Viral Nonstructural Proteins Viral Proteins Viral Tropism Virus Assembly Virus Replication

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