title: Expression and purification of vesicular stomatitis virus N-P complex from Escherichia coli: role in genome RNA transcription and replication in vitro.
abstract: The nucleocapsid protein (N) and phosphoprotein (P) genes of vesicular stomatitis virus (VSV), Indiana serotype, were coexpressed in Escherichia coli BL21(DE3) by using the expression vector pET-3a. The coexpression resulted in the formation of N-P complex. The purified N-P complex was found to inhibit transcription in vitro mediated by viral ribonucleoprotein (RNP) complex in a dose-dependent manner. However, addition of uninfected mammalian cell extracts together with the N-P complex to the transcribing RNP resulted in the synthesis of full-length negative-strand genome RNA. These results indicate that the N-P complex regulated transcription and a cellular factor(s) in combination with the N-P complex may switch the RNA polymerase from transcription to replication mode.
authors.affiliation: Department of Molecular Biology, Research Institute, The Cleveland Clinic Foundation, Ohio 44195, USA.
authors.forename: A K
authors.initials: AK
authors.lastname: Gupta
authors.forename: A K
authors.initials: AK
authors.lastname: Banerjee
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli; D015967:Gene Expression Regulation, Viral; D046911:Macromolecular Substances; D019251:Nucleocapsid / Q000235:genetics*; D019590:Nucleocapsid Proteins; D010750:Phosphoproteins; D011485:Protein Binding; D011994:Recombinant Proteins; D012261:Ribonucleoproteins / Q000378:metabolism; D014158:Transcription, Genetic; D014721:Vesicular stomatitis Indiana virus / Q000737:chemistry / Q000235:genetics*; D015678:Viral Structural Proteins / Q000235:genetics*; D014779:Virus Replication
chemical_list: D046911:Macromolecular Substances; D019590:Nucleocapsid Proteins; C081250:P protein, Vesicular stomatitis virus; D010750:Phosphoproteins; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; D015678:Viral Structural Proteins