Rebinding of transcriptase components (L and NS proteins) to the nucleocapsid template of vesicular stomatitis virus

Mellon, Michael; Emerson, Suzanne U.

doi:10.1128/jvi.27.3.560-567.1978

Cited by 107 publications

(51 citation statements)

References 14 publications

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“…[2][3][4][5] The viral RNA-dependent RNA polymerase consists of two subunits, the large protein (L) that contains the polymerase activity and a polymerase cofactor, the phosphoprotein (P) that binds L to the N-RNA. [6][7][8][9][10][11] The polymerase complex cannot transcribe or replicate the naked vRNA; vRNA has to be bound to N in order to be a functional template. 12 P has a second role in the viral life-cycle as a chaperone for N that is not yet bound to vRNA (N 0 ).…”

Section: Introductionmentioning

confidence: 99%

Structure and Function of the C-terminal Domain of the Polymerase Cofactor of Rabies Virus

Mavrakis

McCarthy

Roche

et al. 2004

Journal of Molecular Biology

103

110

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The phosphoprotein (P) of rabies virus binds the viral polymerase to the nucleoprotein (N)-RNA template for transcription and replication. By limited protease digestion we defined a monomeric C-terminal domain of P that can bind to N-RNA. The atomic structure of this domain was determined and previously described mutations that interfere with binding of P to N-RNA could now be interpreted. There appears to be two features involved in this activity situated at opposite surfaces of the molecule: a positively charged patch and a hydrophobic pocket with an exposed tryptophan side-chain. Other previously published work suggests a conformational change in P when it binds to N-RNA, which may imply the repositioning of two helices that would expose a hydrophobic groove for interaction with N. This domain of rabies virus P is structurally unrelated to the N-RNA binding domains of the phosphoproteins of Sendai and measles virus that are members of the same order of viruses, the non-segmented negative strand RNA viruses. The implications of this finding for the evolution of this virus group are discussed.

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Section: Introductionmentioning

confidence: 99%

Structure and Function of the C-terminal Domain of the Polymerase Cofactor of Rabies Virus

Mavrakis

McCarthy

Roche

et al. 2004

Journal of Molecular Biology

103

110

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“…The enzymes necessary for mRNA synthesis, namely an RNA dependent RNA polymerase (RdRp) and a set of capping enzymes, reside within the viral large protein (L) [17,19,[21][22][23][24][25][26][27]. VSV L protein cannot engage the N-RNA template directly, but instead depends on the viral phoshoprotein (P) to facilitate the interaction [28][29][30][31][32][33]. Messenger RNA polyadenylation is also catalyzed by L through reiterative transcription by the RdRp of a U7 tract that resides at the end of each gene [34][35][36][37][38].…”

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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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“…6). Thus, the NS(NJ) requirement may possibly be necessary to stabilize the heterologous complex (23). Thus, it seems that there may be two distinct sites of interaction of NS(IND) and NS(NJ) on the transcribing N-RNA(IND) complex.…”

Section: Discussionmentioning

confidence: 99%

Specific interactions of vesicular stomatitis virus L and NS proteins with heterologous genome ribonucleoprotein template lead to mRNA synthesis in vitro

Banerjee

1984

J Virol

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Two dissociable proteins, L and NS, and N-RNA template were purified from two serologically distinct vesicular stomatitis viruses, Indiana [VSV(IND)] and New Jersey [VSV(NJ)]. Requirements for RNA synthesis in heterologous reconstitution reactions in vitro were studied. The L and NS proteins of VSV(NJ) failed to synthesize full-length leader RNA and mRNAs in vitro when reconstituted with N-RNA(IND) template. However, when purified homologous NS(IND) was added to the reaction mixture, mRNA synthesis ensued. The requirements for transcription of N-RNA(NJ) template were different from those for N-RNA(IND). For RNA synthesis, transcription specifically required L(NJ), but the NS(NJ) and NS(IND) proteins were interchangeable. This suggests that there are specific domains on the L(NJ) protein, at which NS proteins of both serotypes may interact to form an active RNA polymerase complex, whereas L(IND) lacked such domains for interaction with NS(NJ). The function of the L protein appeared primarily to initiate RNA chains, and the NS protein was required for chain elongation. The results of these in vitro complementation experiments are discussed in light of previous in vivo complementation studies. 628

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Rebinding of transcriptase components (L and NS proteins) to the nucleocapsid template of vesicular stomatitis virus

Cited by 107 publications

References 14 publications

Structure and Function of the C-terminal Domain of the Polymerase Cofactor of Rabies Virus

Structure and Function of the C-terminal Domain of the Polymerase Cofactor of Rabies Virus

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

Specific interactions of vesicular stomatitis virus L and NS proteins with heterologous genome ribonucleoprotein template lead to mRNA synthesis in vitro

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