Rebinding of transcriptase components (L and NS proteins) to the nucleocapsid template of vesicular stomatitis virus.
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. The nucleocapsid-bound L and NS proteins were separated from unbound proteins on a glycerol gradient. The rebinding of L and NS proteins mimics the in vivo binding in that at saturation the ratio of L and NS molecules to N molecules is approximately the same as observed in the intact virion. L and NS proteins were separated and added back independently and in combination to the template. The purified NS protein bound to the template in the absence of L protein. However, the L protein binding appeared to depend on the presence of NS protein. The presence of Mg2+ and nucleotides, which is required for transcription, was not necessary for the rebinding of L and NS proteins.
D002213:Capsid / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D011485:Protein Binding; D012367:RNA, Viral / Q000378:metabolism*; D013698:Templates, Genetic; D014158:Transcription, Genetic; D018116:Vesiculovirus / Q000235:genetics / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism*
D012367:RNA, Viral; D014764:Viral Proteins; D012321:DNA-Directed RNA Polymerases