The vesicular stomatitis virus matrix protein inhibits transcription from the human beta interferon promoter

Abstract

In cells infected by wild-type (wt) vesicular stomatitis virus (VSV) Indiana, host transcription is severely inhibited. DNA cotransfection studies have implicated the VSV matrix (M) protein in this process (B. L. Black and D. S. Lyles, J. Virol. 66:4058-4064, 1992). The M protein inhibited transcription not only from viral promoters in plasmids but also from the chromosomally integrated human immunodeficiency virus type 1 (HIV-1) provirus promoter (S.-Y. Paik, A. C. Banerjea, G. G. Harmison, C.-J. Chen, and M. Schubert, J. Virol. 69:3529-3537, 1995). In this study, we investigated the effect of wt VSV M protein on expression of a reporter gene under control of a cellular promoter (beta-interferon [IFN-␤] promoter), using double transient transfections in BHK and COS-1 cells. The cellular IFN-␤ promoter was as susceptible to the inhibitory effect of the M protein as the viral promoters used previously. Viral proteins N, P, and G had no significant effect on reporter gene expression. The M protein gene from VSV mutant T1026R1, which is defective in host transcription inhibition, was cloned and sequenced, and its effect on reporter gene expression was tested. The mutant M protein had a methionine-to-arginine change at position 51 in the protein sequence and did not inhibit transcription from either the IFN-␤ promoter or viral promoters. This VSV mutant is a good inducer of IFN, as opposed to the wt virus, which suppresses IFN induction. These results show that the M protein inhibits transcription from cellular as well as viral promoters and that the M protein does not regulate the IFN promoter any differently from viral promoters. While the M protein may play a role in IFN gene regulation, other viral or cellular factors that provide specificity to the induction process must also be involved.Houston for quantitating the CAT assays, R. Schloemer for providing the pTWU54 plasmid, M. Schubert for providing the VSV expression vectors, J. Rose and E. Kretzschmar for the VSV polyclonal antibody, and S. Beausoleil and L. Poliquin for the sequence of the M gene from the HR strain of VSV (Indiana).

Interplay between Innate Immunity and Negative-Strand RNA Viruses: towards a Rational Model

“…VOL. 75,2011 INTERPLAY BETWEEN RNA VIRUSES AND INNATE IMMUNITY 471 alternative reading frame (C and Y proteins) or RNA editing (V, D, and W proteins) (144). The virus cycle starts by the binding of viral envelope glycoproteins to a plasma membrane receptor.…”

“…5). This activity is genetically separable from its role in virus structure and assembly as shown by M mutants that are selectively defective (5,22,75,78). When expressed alone by transfection, M protein inhibits transcription by all three host RNA polymerases (3) and the nuclear-cytoplasmic transport of host RNA (102,294).…”

Neurovirulence and Immunogenicity of Attenuated Recombinant Vesicular Stomatitis Viruses in Nonhuman Primates

“…The M protein is the most abundant protein in the virus particle, forming a layer between the lipid envelope and the nucleocapsid core. The M protein also has a major role in particle budding (14,15) and regulation of viral transcription (16,17) and as an agonist of host innate immune responses (18)(19)(20). The virus G protein forms trimeric spike-like projections on the particle surface that enable the virus to dock with a host cell receptor(s).…”

Viral suppression of the interferon system

“…The matrix (M) protein of vesicular stomatitis virus (VSV) is also a potent host cell shutoff factor which inhibits basal transcription [51], impairs nuclear-cytoplasmic transport of RNAs and proteins [52], and inactivates translation factors [53]. As is the case with bunyavirus NSs, the biological significance of VSV M-mediated shutoff is to suppress IFN induction [54,55]. Also, proteinases of Picornaviruses (e.g.…”