Inhibition of vesicular stomatitis virus infection by spike glycoprotein. Evidence for an intracellular, G protein-requiring step.
In an assay measuring virus-directed RNA synthesis, infection of BHK cells by a standard test dose of vesicular stomatitis virus (VSV) was inhibited by ultraviolet light-irradiated wt VSV and by ts 045, one of a number of thermolabile, temperature-sensitive G protein mutants of VSV. After heat treatment for 1 h at 45 degrees C, the thermolabile mutants were no longer able to inhibit the VSV infection. In contrast, the thermolabile M protein mutant ts G31 and the nonthermolabile G protein mutant ts 044 could still inhibit the test VSV dose. Thus, the presence of G protein in its native conformation was necessary for inhibition of infection. There was little difference in the binding to cells or the internalization to a trypsin-resistant state of ts 045 or wt VSV before and after heat treatment, and there was no evidence of specific saturable receptors on the cell surface. None of the irradiated virions at concentrations that gave maximal inhibition of infection could prevent binding of infectious VSV to, or internalization by, BHK cells. The G protein-specific inhibition, therefore, did not occur at the cell surface but must have occurred at some intracellular site, which has been suggested to be the lysome. The lysosomal inhibitor chloroquine, when added with the infecting virus, completely inhibited VSV infection at all multiplicities of infection tested, and it gave 50% inhibition when added to 1.5 h after infection. The possible importance of the lysosome in the intracellular pathway of infection is discussed.
D000818:Animals; D002460:Cell Line; D002738:Chloroquine / Q000494:pharmacology; D006224:Cricetinae; D006023:Glycoproteins / Q000235:genetics / Q000494:pharmacology*; D007668:Kidney; D009154:Mutation; D012334:RNA, Neoplasm / Q000096:biosynthesis; D011991:Receptors, Virus / Q000378:metabolism; D013696:Temperature; D014466:Ultraviolet Rays; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000494:pharmacology*; D014779:Virus Replication / Q000187:drug effects*
D006023:Glycoproteins; D012334:RNA, Neoplasm; D011991:Receptors, Virus; D014764:Viral Proteins; D002738:Chloroquine