Effect of temperature-sensitive mutations on the virion-associated RNA transcriptase of vesicular stomatitis virus

Szilágyi, J. F.; Pringle, C. R.

doi:10.1016/0022-2836(72)90351-8

Cited by 67 publications

(60 citation statements)

References 11 publications

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“…These data are summarized in Table 1. We assayed this virus at 31 and 39°C due to the original temperature-sensitive phenotype described for ts(G)16 (33,39). We found that the F1488S rVSV had a slight reduction in RNA transcription levels at 39°C and that the plaque size was reduced at 39°C compared to 31°C.…”

Section: Analysis Of Poly(a) Tail Lengthmentioning

confidence: 99%

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S -Adenosyl Homocysteine-Induced Hyperpolyadenylation of Vesicular Stomatitis Virus mRNA Requires the Methyltransferase Activity of L Protein

Galloway

Wertz

2008

J Virol

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There are many unique aspects of vesicular stomatitis virus (VSV) transcription. In addition to its unusual mRNA capping and methyltransferase mechanisms, the addition of S-adenosyl homocysteine (SAH), which is the by-product and competitive inhibitor of S-adenosyl methionine (SAM)-mediated methyltransferase reactions, leads to synthesis of poly(A) tails on the 3 end of VSV mRNAs that are 10-or 20-fold longer than normal. The mechanism by which this occurs is not understood, since it has been shown that productive transcription is not dependent on 5 cap methylation and full-length VSV mRNAs can be synthesized in the absence of SAM. To investigate this unusual phenotype, we assayed the effects of SAH on transcription using a panel of recombinant viruses that contained mutations in domain VI of the VSV L protein. The L proteins we investigated displayed a range of 5 cap methyltransferase activities. In the present study, we show that the ability of the VSV L protein to catalyze methyl transfer correlates with its sensitivity to SAH with respect to polyadenylation, thereby indicating an intriguing connection between 5 and 3 end mRNA modifications. We also identified an L protein mutant that hyperpolyadenylates mRNA irrespective of the presence or absence of exogenous SAH. Further, the data presented here show that the wild-type L protein hyperpolyadenylates a percentage of VSV mRNAs in infected cells as well as in vitro. Vesicular stomatitis virus (VSV) is a member of the Mononegavirales.Viruses within this order have a single strand of nonsegmented negative-sense genomic RNA. For VSV, there are five genes, and the order is 3Ј-(leader)-N-P-M-G-L-(trailer)-5Ј (3). The nucleocapsid (N) protein encapsidates the viral RNA genome and the N:RNA complex comprises the active template for all viral RNA synthesis. The phosphoprotein (P) is a cofactor for the RNA-dependent RNA polymerase (RdRp) and a solubility factor for the N protein to prevent N-protein aggregation (15,19). The matrix (M) is the most abundant structural protein and is responsible for the bullet-shaped morphology of the virion (30). The glycoprotein (G) provides both attachment and fusion functions for entry of the virus into the host cell (31). The large (L) protein is the catalytic component of the viral RdRp (11). Expression of the genes of VSV is controlled primarily at the level of transcription (reviewed in reference 7). The general mechanism for transcription of the Mononegavirales is that the RdRp enters the genome at a single 3Ј entry site and transcribes each gene in an obligatorily sequential manner (1-3). A central feature of this mode of transcription is that RdRp access to a downstream gene for transcription is dependent upon prior termination of the upstream mRNA. Due to a poorly understood process, known as attenuation, which has been localized to each gene junction, there is a gradient of mRNA synthesis, such that the levels of each mRNA decrease with each successive RdRp initiation event:genes closer to the 3Ј entry site are transcribed mo...

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Section: Analysis Of Poly(a) Tail Lengthmentioning

confidence: 99%

“…ts(G)16 was isolated in 1970 based on its ability to grow at 31°C but not at 39°C (39). The mRNAs synthesized by the ts(G)16 polymerase were excessively polyadenylated at the 3Ј end (21).…”

Section: Vesicular Stomatitis Virus (Vsv) Is a Member Of The Mononegamentioning

confidence: 99%

S -Adenosyl Homocysteine-Induced Hyperpolyadenylation of Vesicular Stomatitis Virus mRNA Requires the Methyltransferase Activity of L Protein

Galloway

Wertz

2008

J Virol

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“…By heteroduplex analysis we can assign complementation groups III and IV to genes coding for the M and N proteins respectively. Complementation group I was not examined because the protein results are clear-cut (Szilagyi & Pringle, 1972;Hunt et al, 1976). The inability to assign complementation groups II and V to a single gene by this method can be explained in several ways.…”

Section: Discussionmentioning

confidence: 99%

“…FREEMAN AND A. S. HUANG the polymerase gene, which codes for the largest (L) of the VSV proteins (Szilagyi & Pringle, 1972;Hunt etal., 1976).…”

Section: Introductionmentioning

confidence: 99%

Mapping Temperature-sensitive Mutants of Vesicular Stomatitis Virus by RNA Heteroduplex Formation

Freeman

Huang

1981

Journal of General Virology

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SUMMARYDuplex RNA molecules made by hybridization of virion and mRNA of vesicular stomatitis virus (VSV) were digested with ribonuclease and separated into five size classes, each containing the gene and the mRNA for one of the VSV proteins. Denaturation of the duplexes yielded full size mRNA lacking poly(A) tails. Utilizing duplex formation between the RNAs from VSV temperature-sensitive (ts) mutants and their revertants and subsequent RNase digestion under varying salt conditions, specific cleavages within a certain duplex were seen for representative mutants from complementation groups III, IV and V. Specific cleavages were not seen for a group II mutant. From these results gene assignments cannot be made for group II; equivocal assignments are made for group III and clear assignments made for group IV and V. The assignment for the group V mutants, however, does not conform to expectations. Nevertheless, from these studies and other published ones, there is the suggestion that interactions may exist between the gene products of complementation groups II and V during VSV transcription and morphogenesis. These results also support the lack of transcriptional splicing for VSV mRNAs.

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“…These should probably be less virulent, but we have not yet determined the LDs0 of these mutants in mice. Presumably the rate of generation/amplification of DI particles would be regulated by viral replication/encapsidation genes (Leppert et al, 1977;Huang, 1977) and mutants of viral complementation group I (L protein polymerase gene) are the most abundant mutants of VSV (Pringle, 1970(Pringle, , 1977Szilagyi & Pringle, 1979). Some RNA viruses might generate DI particles extremely rapidly even from common wild-type and mutant strains.…”

Section: Short Communicationmentioning

confidence: 99%

Very Rapid Generation/Amplification of Defective Interfering Particles by Vesicular Stomatitis Virus Variants Isolated from Persistent Infection

DePolo

Holland

1986

Journal of General Virology

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SUMMARYMultiply cloned variants of vesicular stomatitis virus (VSV) were found to generate/amplify defective interfering (DI) particles at a rate greatly exceeding the rates normally observed for wild-type VSV (or for other mutants of VSV). A single undiluted passage of the first clonal pool of this variant virus produced concentrated visible bands of DI particles on sucrose gradients whereas wild-type and other mutant strains of VSV required from three to six or more serial undiluted passages. Since DI particle amplication by wild-type VSV at each undiluted passage can exceed 10000-fold enrichment, these variant virus clones were generating/amplifying DI particles many millions of times more rapidly than were wild-type and other mutant strains of VSV. This rate of generation/amplification is so high that it was not feasible to obtain accurate estimates of the rates of generation (or amplification) of these DI particles. Stampfer et al. (1971) first demonstrated that defective interfering (DI) particles can be eliminated from vesicular stomatitis virus (VSV) stocks by cloning, preferably multiple cloning, of infectious virus. Holland et al. (1976 a) demonstrated that even the first clonal pools prepared from multiply cloned virus are already contaminated with seed quantities of newly generated DI particles, and this was confirmed for both VSV and Sendai virus DI particles

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Effect of temperature-sensitive mutations on the virion-associated RNA transcriptase of vesicular stomatitis virus

Cited by 67 publications

References 11 publications

S -Adenosyl Homocysteine-Induced Hyperpolyadenylation of Vesicular Stomatitis Virus mRNA Requires the Methyltransferase Activity of L Protein

S -Adenosyl Homocysteine-Induced Hyperpolyadenylation of Vesicular Stomatitis Virus mRNA Requires the Methyltransferase Activity of L Protein

Mapping Temperature-sensitive Mutants of Vesicular Stomatitis Virus by RNA Heteroduplex Formation

Very Rapid Generation/Amplification of Defective Interfering Particles by Vesicular Stomatitis Virus Variants Isolated from Persistent Infection

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