Dissociation and Reconstitution of the Transcriptase and Template Activities of Vesicular Stomatitis B and T Virions

Emerson, Suzanne U.; Wagner, Robert R.

doi:10.1128/jvi.10.2.297-309.1972

Cited by 218 publications

(115 citation statements)

References 18 publications

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“…Sindbis virus-induced interference does not appear to affect host cell activation of the VSV transcriptase. Although the mechanism of activation of this enzyme is not understood, it probably involves dissolution of' the membrane enveloping the VSV nucleocapsid, since in vitro activation is brought about with carefully controlled detergent concentrations (2) and the transcriptase activity has been shown to purify with the proteins of the nucleocapsid rather than with those of the envelope (4,8,27).…”

Section: Characterization Of Interference Inductionmentioning

confidence: 99%

Mechanism of Sindbis Virus-Induced Intrinsic Interference with Vesicular Stomatitis Virus Replication

Hunt

Marcus

1974

J Virol

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Heterologous viral interference is induced by Sindbis virus against vesicular stomatitis virus (VSV) in a manner analogous to intrinsic interference with Newcastle disease virus replication. Interference in both systems (i) depends upon early expression of the inducing virus genome, (ii) shows similar kinetics of induction, (iii) does not involve interferon action, and (iv) appears to be manifest as an all-or-none effect. VSV can be added to the list of viruses blocked by intrinsic interference. Sindbis virus-induced intrinsic interference with VSV replication is not mediated through homotypic interference by defective interfering particles; rather, T-particle and B-particle synthesis is inhibited. Significantly, intrinsic interference has no effect on primary transcription directed by the virion-associated transcriptase in VSV-challenged cells. However, Sindbis virus appears to induce interference with the VSV-RNA synthesized subsequent to primary transcription, namely, that which is dependent on protein synthesis. Thus, the target of intrinsic interference appears to be a reaction subsequent to primary transcription but prior to the appearance of protein synthesis-dependent VSV RNA, secondary transcription.Virion-associated RNA transcriptases are presumably the first viral enzymes to function in the replication of negative-strand viruses (1). In addition, development of an interferonmediated antiviral state in cells correlates quantitatively with the inhibition of vesicular stomatitis virus (VSV) virion transcriptase activity (13, 15). The virion transcriptase of VSV thus appears to be a target of interferonmediated interference with replication of this virus. In contrast, interference with VSV replication by defective interfering particles of VSV, homotypic interference, does not affect the synthesis of VSV RNA by the virion-associated transcriptase (11). We have characterized, and describe herein, yet a third type of interference in which VSV transcriptase activity can be measured, viz., interference with VSV replication induced by Sindbis virus. Our results show that this heterotypic interference system is analogous to that described by Marcus and Carver (14) as intrinsic interference.In the absence of an assay for cells refractory to infection by VSV comparable to that used in the hemadsorption-negative plaque or cell test for viruses inducing intrinsic interference against Newcastle disease virus (NDV), inter-'Present address:

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Section: Characterization Of Interference Inductionmentioning

confidence: 99%

Mechanism of Sindbis Virus-Induced Intrinsic Interference with Vesicular Stomatitis Virus Replication

Hunt

Marcus

1974

J Virol

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“…As VSV RNA synthesis occurs entirely within the cytoplasm, viral RNA synthesis is not subject to the inhibitory effects of M on RNA polymerase II and mRNA export from the nucleus. The VSV RNA synthesis machinery comprises a ribonucleoprotein complex of the negative-sense genomic RNA completely encased by a nucleocapsid protein (N) sheath and associated with the viral polymerase complex [13]. The viral transcriptase copies the N-RNA template into 5 monocistronic mRNAs that are structurally indistinct to those of the host-cell with respect to their 5 0 cap and 3 0 polyadenylate tail [14][15][16][17][18][19][20].…”

mentioning

confidence: 99%

Global analysis of polysome-associated mRNA in vesicular stomatitis virus infected cells

Neidermyer¹,

Whelan²

2019

PLoS Pathog

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Infection of mammalian cells with vesicular stomatitis virus (VSV) results in the inhibition of cellular translation while viral translation proceeds efficiently. VSV RNA synthesis occurs entirely within the cytoplasm, where during transcription the viral polymerase produces 5 mRNAs that are structurally indistinct to cellular mRNAs with respect to their 5′ cap-structure and 3′-polyadenylate tail. Using the global approach of massively parallel sequencing of total cytoplasmic, monosome- and polysome-associated mRNA, we interrogate the impact of VSV infection of HeLa cells on translation. Analysis of sequence reads in the different fractions shows >60% of total cytoplasmic and polysome-associated reads map to the 5 viral genes by 6 hours post-infection, a time point at which robust host cell translational shut-off is observed. Consistent with an overwhelming abundance of viral mRNA in the polysome fraction, the reads mapping to cellular genes were reduced. The cellular mRNAs that remain most polysome-associated following infection had longer half-lives, were typically larger, and were more AU rich, features that are shared with the viral mRNAs. Several of those mRNAs encode proteins known to positively affect viral replication, and using chemical inhibition and siRNA depletion we confirm that the host chaperone heat shock protein 90 (hsp90) and eukaryotic translation initiation factor 3A (eIF3A)—encoded by 2 such mRNAs—support viral replication. Correspondingly, regulated in development and DNA damage 1 (Redd1) encoded by a host mRNA with reduced polysome association inhibits viral infection. These data underscore the importance of viral mRNA abundance in the shut-off of host translation in VSV infected cells and link the differential translatability of some cellular mRNAs with pro- or antiviral function.

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“…The VSV particles are bullet-shaped (ϳ80 nm by ϳ200 nm) and comprise a ribonucleoprotein core of genomic RNA and N (nucleocapsid) protein surrounded by a matrix (M) protein layer, enveloped by the host cell plasma membrane containing the transmembrane viral glycoprotein (G) (12). The phosphoprotein (P) and large (L) polymerase protein associate to form the functional viral RNA polymerase, which performs both mRNA transcription and genome replication from the nucleocapsid template (13,14). Viral mRNA transcription initiates at the 3= end of the genome and proceeds with a pronounced 3=-to-5= gradient of gene expression, leading to abundant N protein and successively decreasing levels of P, M, G, and L proteins in the infected cell, providing virus proteins an optimal ratio for subsequent viral genome replication and the assembly of mature virus particles (11,15).…”

mentioning

confidence: 99%

Recombinant Isfahan Virus and Vesicular Stomatitis Virus Vaccine Vectors Provide Durable, Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge

Nasar

Matassov

Seymour

et al. 2017

J Virol

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The demonstrated clinical efficacy of a recombinant vesicular stomatitis virus (rVSV) vaccine vector has stimulated the investigation of additional serologically distinct Vesiculovirus vectors as therapeutic and/or prophylactic vaccine vectors to combat emerging viral diseases. Among these viral threats are the encephalitic alphaviruses Venezuelan equine encephalitis virus (VEEV) and Eastern equine encephalitis virus (EEEV), which have demonstrated potential for natural disease outbreaks, yet no licensed vaccines are available in the event of an epidemic. Here we report the rescue of recombinant Isfahan virus (rISFV) from genomic cDNA as a potential new vaccine vector platform. The rISFV genome was modified to attenuate virulence and express the VEEV and EEEV E2/E1 surface glycoproteins as vaccine antigens. A single dose of the rISFV vaccine vectors elicited neutralizing antibody responses and protected mice from lethal VEEV and EEEV challenges at 1 month postvaccination as well as lethal VEEV challenge at 8 months postvaccination. A mixture of rISFV vectors expressing the VEEV and EEEV E2/E1 glycoproteins also provided durable, single-dose protection from lethal VEEV and EEEV challenges, demonstrating the potential for a multivalent vaccine formulation. These findings were paralleled in studies with an attenuated form of rVSV expressing the VEEV E2/E1 glycoproteins. Both the rVSV and rISFV vectors were attenuated by using an approach that has demonstrated safety in human trials of an rVSV/HIV-1 vaccine. Vaccines based on either of these vaccine vector platforms may present a safe and effective approach to prevent alphavirus-induced disease in humans.IMPORTANCE This work introduces rISFV as a novel vaccine vector platform that is serologically distinct and phylogenetically distant from VSV. The rISFV vector has been attenuated by an approach used for an rVSV vector that has demonstrated safety in clinical studies. The vaccine potential of the rISFV vector was investigated in a well-established alphavirus disease model. The findings indicate the feasibility of producing a safe, efficacious, multivalent vaccine against the encephalitic alphaviruses VEEV and EEEV, both of which can cause fatal disease. This work also demonstrates the efficacy of an attenuated rVSV vector that has already demonstrated safety and immunogenicity in multiple HIV-1 phase I clinical studies. The absence of serological cross-reactivity between rVSV and rISFV and their phylogenetic divergence within the Vesiculovirus genus indicate potential for two stand-alone vaccine

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Dissociation and Reconstitution of the Transcriptase and Template Activities of Vesicular Stomatitis B and T Virions

Cited by 218 publications

References 18 publications

Mechanism of Sindbis Virus-Induced Intrinsic Interference with Vesicular Stomatitis Virus Replication

Mechanism of Sindbis Virus-Induced Intrinsic Interference with Vesicular Stomatitis Virus Replication

Global analysis of polysome-associated mRNA in vesicular stomatitis virus infected cells

Recombinant Isfahan Virus and Vesicular Stomatitis Virus Vaccine Vectors Provide Durable, Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge

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