Vesicular stomatitis virion-associated transcriptase activity was suppressed in vitro by a synthetic 21 amino acid oligopeptide prepared to mimic the carboxy-terminus of NS protein.
To study the biological function of the NS protein of vesicular stomatitis virus (VSV), we prepared 21 species of synthetic oligopeptides with 11-21 amino acid residues, corresponding to every portion of the amino acid sequence of NS protein (Indiana serotype), and tested their effects on the VS virion (VSV) transcriptase activity in vitro. Only one peptide affected the virion-associated transcriptase activity of VSV Indiana, by reducing the incorporation of [3H]GMP into acid-insoluble fraction (IC50 = 26 microM). This peptide, the amino acid sequence of which corresponded to the carboxy (C)-terminal region of NS protein, also inhibited the New Jersey serotype virus transcriptase activity, as expected from a high degree of homology found between the amino acid sequences of the C-terminal regions of NS protein of both serotype viruses. Electrophoretic analysis on acrylamide gels of RNA transcripts revealed that the inhibitory synthetic peptide decreased the frequency of the initiation of transcription with no apparent effect on the chain-elongation process of viral transcription. As expected from its highly conserved amino acid sequence, these results suggest that the C-terminal domain of VSV NS protein is involved in initiating viral RNA synthesis.
Yamashita
T
T
Department of Molecular Microbiology, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Kawai
A
A
D000595:Amino Acid Sequence; D000818:Animals; D002213:Capsid / Q000494:pharmacology*; D002478:Cells, Cultured; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000494:pharmacology*; D012367:RNA, Viral / Q000096:biosynthesis*; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects / Q000201:enzymology*; D018116:Vesiculovirus; D014758:Viral Core Proteins / Q000494:pharmacology*; D017361:Viral Nonstructural Proteins
D009842:Oligopeptides; D012367:RNA, Viral; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins; D012321:DNA-Directed RNA Polymerases