Location of the binding domains for the RNA polymerase L and the ribonucleocapsid template within different halves of the NS phosphoprotein of vesicular stomatitis virus.

AUTOR(ES)

Emerson, S U

RESUMO

Recombinant DNA techniques were used to delete regions of a cDNA clone of the phosphoprotein NS gene of vesicular stomatitis virus. 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. Run-off transcripts were synthesized and translated in vitro to provide [35S]methionine-labeled complete NS or deletion mutant NS proteins. Immune coprecipitation assays involving these proteins were developed to map the regions of the NS protein responsible for binding to the structural viral nucleocapsid protein N and the catalytic RNA polymerase protein L. The data indicate the NS protein is a bivalent protein consisting of two discrete functional domains. Contrary to previous suggestions, the negatively charged amino-terminal half of NS protein binds to L protein, while the carboxyl-terminal half of NS protein binds to both soluble recombinant nucleocapsid protein N and viral ribonucleocapsid template.

Documentos Relacionados

• Phosphorylation sites on phosphoprotein NS of vesicular stomatitis virus.
• Cooperative binding of multimeric phosphoprotein (P) of vesicular stomatitis virus to polymerase (L) and template: pathways of assembly.
• Rebinding of transcriptase components (L and NS proteins) to the nucleocapsid template of vesicular stomatitis virus.
• Both NS and L proteins are required for in vitro RNA synthesis by vesicular stomatitis virus.
• RNA polymerase-associated interactions near template promoter sequences of defective interfering particles of vesicular stomatitis virus.