Disease relevance of VSIVgp2

• Structure of the gene N:gene NS intercistronic junction in the genome of vesicular stomatitis virus [1].
• The results of both the in vivo and in vitro experiments show that the replication of the VSV genome is controlled by the availability of the nucleocapsid protein, even when the polymerase has access to the host factors and multiple phosphorylated forms of the NS protein thought to be involved in genomic RNA synthesis [2].
• The complete NS gene and four mutant genes containing internal or terminal deletions were inserted into a modified pGem4 vector under the transcriptional control of the phage T7 promoter [3].
• The activity, together with the viral L and NS proteins, is removed from virions by treatment with 0.8 M NaCl [4].
• Upon infection of Chinese hamster ovary cells (CHO), vesicular stomatitis (VSV) virus synthesizes two membrane proteins (the VSV glycoprotein and the VSV matrix or membrane (M) protein) and three nonmembrane proteins (the VSV nucleocapsid, the viral transcriptase, and an NS protein) [5].

High impact information on VSIVgp2

• Phosphorylation of NS by the L protein was abrogated by substitution of either serine 236 or serine 242 [6].
• We have investigated the functional significance of phosphoserine residues that lie in the L protein-binding domain between amino acids 213 and 247 of the phosphoprotein (NS) of vesicular stomatitis virus [6].
• NS consists of a highly acidic NH2-terminal domain and a basic COOH-terminal domain [7].
• These observations indicate that NS probably functions as an activator protein in which the acidic domain stimulates transcription of the viral genes by interacting with the RNA polymerase as observed for eukaryotic cellular transcription activators [7].
• Transcription of the cDNA in vitro resulted in the synthesis of NS mRNA, which was subsequently translated into NS protein in a cell-free rabbit reticulocyte system [8].

Chemical compound and disease context of VSIVgp2

• NS protein of vesicular stomatitis virus was shown to migrate with a mobility consistent with the molecular weight predicted from the published cDNA sequence on polyacrylamide gels containing the detergent cetyltrimethylammonium bromide at low pH [9].
• The phosphoprotein NS of vesicular stomatitis virus which accumulates within the infected cell cytoplasm is phosphorylated at multiple serine and threonine residues (G. M. Clinton and A. S. Huang, Virology 108:510-514, 1981; Hsu et al., J. Virol. 43:104-112, 1982) [10].
• The L and NS proteins of vesicular stomatitis virions (New Jersey serotype) were solubilized with Triton X-100 and high-salt buffer and recombined with purified nucleocapsids under conditions similar to those used to reconstitute transcriptase activity in vitro [11].
• Peptide maps, obtained by limited proteolysis of 32P-labeled wild-type and mutant NS proteins with Staphylococcus aureus V8 protease and papain, revealed striking differences which suggested that the mutant alteration could involve an aspartic or glutamic acid residue [12].
• Treatment of nucleocapsids with 5'-p-fluorosulfonylbenzoyl adenosine resulted in greater than 90% inhibition of NS phosphorylation but had no effect on RNA polymerase activity [13].

Biological context of VSIVgp2

• Nucleotide sequences of the mRNA's encoding the vesicular stomatitis virus N and NS proteins [14].
• Instead, the initiation of transcription is more likely influenced by the position of the polymerase binding site relative to the 3' end or by requisite interactions between the catalytic polymerase component (L) and the proposed initiator protein (NS) [15].
• Methylation protection studies suggested that the NS protein component of the RNA polymerase of vesicular stomatitis virus contacts the RNA templates of defective interfering (DI) particles at the sequence 3'...GUCUAUUUUUAUUUUUGGUG...5',17 to 37 nucleotides downstream from the site of initiation of in vitro transcription [15].
• These results indicate that phosphorylation of serines 236 and 242 in the NS protein regulates its binding with the L protein and the N-RNA template and is essential for activation of viral RNA synthesis [6].
• These data are consistent with the idea that the reversion of the temperature-sensitive phenotype of tsE1 can be accompanied by production of a significantly larger NS protein [16].

Anatomical context of VSIVgp2

• Our results suggest that the VSV glycoprotein is synthesized exclusively on membrane polyribosomes, while at least some of the M, N, and NS proteins are made on free polyribosomes [5].
• In contrast, the viral proteins NS and M appeared to be selectively degraded in NK cell extracts [17].
• Oligodeoxyribonucleoside methylphosphonates which have sequences complementary to the initiation codon regions of N, NS, and G vesicular stomatitis virus (VSV) mRNAs were tested for their ability to inhibit translation of VSV mRNA in a cell-free system and in VSV-infected mouse L cells [18].
• The NS gene encodes a protein of 265 amino acids which was expressed from a simian virus 40 vector in COS cells [19].
• It was further observed that RNP stripped of L protein but containing residual NS protein was capable of initiating virus production or protein synthesis when microinjected in vero cells or in oocytes, respectively [20].

Associations of VSIVgp2 with chemical compounds

• Deficiency of sodium dodecyl sulfate binding to a large negatively charged domain in the NS protein could explain this anomalous electrophoretic mobility [14].
• The VSV and mrna encoding the NS protein is 815 nucleotides long, excluding polyadenylic acid, and encodes a protein of 222 amino acids [14].
• Cyanogen bromide cleavage of NS protein produced a large acidic amino-terminal peptide, as predicted by the sequence, which contained the majority of the phosphate residues [9].
• Although the packaged wild-type and mutant NS proteins were similarly phosphorylated, the mutant NS protein migrated faster than the wild-type NS protein in polyacrylamide slab gels electrophoresed in the presence of sodium dodecyl sulfate [12].
• The nucleocapsid-bound L and NS proteins were separated from unbound proteins on a glycerol gradient [11].

Analytical, diagnostic and therapeutic context of VSIVgp2

• However, chymotryptic peptide mapping has indicated that all forms of NS share a common cluster of phosphorylated residues [21].
• Two-dimensional gel electrophoresis demonstrated that each of the NS proteins consisted of several species, but the isoelectric points of the proteins from different viruses overlapped [16].
• 6. Through immunoprecipitation analysis of the viral soluble protein pool, we found that a complex that usually exists between the N and NS proteins at pH 7.4 was altered in extracts from infected cells maintained at pH 6.6, and this was responsible for the observed effects on viral replication [22].
• This antiserum gave specific reactions of identity by agar immunodiffusion with both cytoplasmic and virion NS protein [23].
• NS phosphoprotein of vesicular stomatitis virus: subspecies separated by electrophoresis and isoelectric focusing [24].

References