Determination of molar ratios of vesicular stomatitis virus induced RNA species in BHK<sub>21</sub>cells

Villarreal, Luis P.; Breindl, Michael; Holland, John J.

doi:10.1021/bi00653a012

Cited by 130 publications

(91 citation statements)

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“…There are conserved, untranslated, intergenic junctions between each gene, which contain the transcription termination and polyadenylation signals for the upstream gene and the transcription reinitiation signals for the subsequent downstream gene (Rose, 1980;Rose & Schubert, 1987;Schnell et al, 1996b). In terms of transcriptional attenuation of the VSV genome, the quantity of transcripts produced from each of the downstream genes is approximately 30 % lower than those of the gene located immediately upstream (Villarreal et al, 1976).…”

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confidence: 98%

Expression and processing of human immunodeficiency virus type 1 gp160 using the vesicular stomatitis virus New Jersey serotype vector system

Kim

Kang

2009

Journal of General Virology

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The Indiana serotype of vesicular stomatitis virus (VSVIND), but not the New Jersey serotype (VSVNJ), has been widely used as a gene expression vector. In terms of prime–boost-based vaccine strategies, it would be desirable to use two different VSV serotypes to avoid immunity against the priming viral vector. Here, we report that we have applied the VSVNJ vector system for expression of the env gene of human immunodeficiency virus type 1 (HIV-1). The HIV-1 env gene was inserted into the VSVNJ vector system at two different sites: between the P and M genes (NP-gp160-MGL) and between the G and L genes (NPMG-gp160-L). The HIV-1 env gene product, gp160, was efficiently expressed and processed in cells infected with either of these two recombinant VSV–HIV-1gp160 viruses. In this study, we have investigated the applicability of the VSVNJ vector system for foreign gene expression.

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confidence: 98%

Expression and processing of human immunodeficiency virus type 1 gp160 using the vesicular stomatitis virus New Jersey serotype vector system

Kim

Kang

2009

Journal of General Virology

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“…Due to transcriptional attenuation at each gene junction, the amount of mRNA transcribed is a function of gene position relative to the promoter. Promoter-proximal genes are transcribed at higher levels than more distal genes (23,54).…”

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Vesicular Stomatitis Virus Glycoprotein Is a Determinant of Pathogenesis in Swine, a Natural Host

Martı́nez

Rodrı́guez

Jiménez

et al. 2003

J Virol

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There are two major serotypes of vesicular stomatitis virus (VSV), Indiana (VSIV) and New Jersey (VSNJV). We recovered recombinant VSIVs from engineered cDNAs that contained either (i) one copy of the VSIV G gene (VSIV-GI); (ii) two copies of the G gene, one from each serotype (VSIV-GNJGI); or (iii) a single copy of the GNJ gene instead of the GI gene (VSIV-GNJ). The recombinant viruses expressed the appropriate glycoproteins, incorporated them into virions, and were neutralized by antibodies specific for VSIV (VSIV-GI), VSNJV (VSIV-GNJ), or both (VSIV-GNJGI), according to the glycoprotein(s) they expressed. All recombinant viruses grew to similar titers in cell culture. In mice, VSIV-GNJ and VSIV-GNJGI were attenuated. However, in swine, a natural host for VSV, the GNJ glycoprotein-containing viruses caused more severe lesions and replicated to higher titers than the parental virus, VSIV-GI. These observations implicate the glycoprotein as a determinant of VSV virulence in a natural host and emphasize the differences in VSV pathogenesis between mice and swine

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“…Spots 1, 3, and 4 were obtained when N protein mRNA was used, whereas spot 6 was the only sequence derived from the G protein mRNA. Spots 2,5,8,9, and 10 were all derived from the mixture of NS and M mRNAs, which do not separate on formamide/polyacrylamide gels (6,7). The source of spot 7 was not identified.…”

Section: Methodsmentioning

confidence: 99%

“…These mRNAs are translated with equal efficiencies in vivo (9). I report here a procedure for obtaining translation initiation sites from the VSV mRNAs synthesized both in VSV-infected cells and in vitro by the VSV virion-associated RNA polymerase (10,11).…”

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Nucleotide sequences of ribosome recgonition sites in messenger RNAs of vesicular stomatitis virus.

Rose¹

1977

Proc. Natl. Acad. Sci. U.S.A.

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The ribosome recognition sites from vesicular stomatitis virus mRNAs have been isolated by ribosome yrotection of the mixture of viral mRNAs followed by comp ete separation of the protected sites by two-dimensional gel electrophoresis. The sites from the four major mRNAs each contain a single AUG initiation codon, indicating that they are true initiation sites, and the complete sequences of the N, NS, and G mRNA sites are reported. The sites do not all share common features nor do they have significant regions complementary to the known sequence at the 3'-terminus of 18S rRNA. The N and NS mRNAs show a nine-nucleotide homology in the first eleven nucleotides extending from the capped 5'-end.In contrast to prokaryotic mRNAs which commonly have multiple translation initiation sites, all eukaryotic mRNAs so far examined have only single active translation initiation sites, suggesting a fundamental difference between eukaryotic and prokaryotic mRNA translation (1). Also, prokaryotic mRNAs lack the capped and methylated 5'-termini that are found on most eukaryotic mRNAs (2). Nucleotide sequences from translation initiation sites in many mRNAs from prokaryotes have been determined in the past few years (see review in ref.3). Comparison of these sequences has suggested a model for initiation site selection involving base pairing between the 3'-end of the RNA from the small ribosome subunit with a sequence in the mRNA (4). By analogy with prokaryotic systems, structural studies on initiation sites are likely to prove valuable in elucidating the mechanisms involved in selection of translation initiation sites on eukaryotic mRNA.Vesicular stomatitis virus (VSV) is an animal virus that produces five well-characterized mRNAs that are complementary to the single-stranded RNA genome (5-8). These mRNAs are translated with equal efficiencies in vivo (9). I report here a procedure for obtaining translation initiation sites from the VSV mRNAs synthesized both in VSV-infected cells and in vitro by the VSV virion-associated RNA polymerase (10, 11). The key feature of the method is the use of unfractionated mRNAs in large-scale preparation of mRNA fragments that are protected from nuclease digestion in a ribosome-mRNA initiation complex. The protected sites are then completely separated from each other by exploiting differences in base composition and size on the two-dimensional gel electrophoretic system of De Wachter and Fiers (12). MATERIALS AND METHODSSynthesis and Ribosome Binding of mRNA. In vivo 32p-labeling of VSV mRNA and periodate oxidation and f3-elimination to remove 7-methylguanosine were as described (13).In vitro syntheses (0.5-ml reaction mixtures) were with individual [a-32P]nucleoside triphosphates at final specific radioactivities of 12.5 Ci/mmol and concentrations of 80 ,uM for each of the labeled triphosphates and 1 mM for the three unlabeled triphosphates (11). Approximately 25 ,ug of poly(A)-containing product mRNA (108 cpm) was obtained from each reaction. Isotopes were purchased from New England Nuc...

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Determination of molar ratios of vesicular stomatitis virus induced RNA species in BHK₂₁cells

Cited by 130 publications

References 15 publications

Expression and processing of human immunodeficiency virus type 1 gp160 using the vesicular stomatitis virus New Jersey serotype vector system

Expression and processing of human immunodeficiency virus type 1 gp160 using the vesicular stomatitis virus New Jersey serotype vector system

Vesicular Stomatitis Virus Glycoprotein Is a Determinant of Pathogenesis in Swine, a Natural Host

Nucleotide sequences of ribosome recgonition sites in messenger RNAs of vesicular stomatitis virus.

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Determination of molar ratios of vesicular stomatitis virus induced RNA species in BHK21cells

Cited by 130 publications

References 15 publications

Expression and processing of human immunodeficiency virus type 1 gp160 using the vesicular stomatitis virus New Jersey serotype vector system

Expression and processing of human immunodeficiency virus type 1 gp160 using the vesicular stomatitis virus New Jersey serotype vector system

Vesicular Stomatitis Virus Glycoprotein Is a Determinant of Pathogenesis in Swine, a Natural Host

Nucleotide sequences of ribosome recgonition sites in messenger RNAs of vesicular stomatitis virus.

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Determination of molar ratios of vesicular stomatitis virus induced RNA species in BHK₂₁cells