Proteins of vesicular stomatitis virus: kinetics and cellular sites of synthesis.
FIVE VIRAL PEPTIDES SYNTHESIZED IN L CELLS INFECTED WITH VESICULAR STOMATITIS (VS) VIRUS WERE IDENTIFIED BY POLYACRYLAMIDE GEL ELECTROPHORESIS AND DESIGNATED AS FOLLOWS: nucleoprotein N, a membrane glycoprotein G, a membrane surface protein S, and two nonstructural proteins NS1 and NS2. A slowly migrating minor structural protein L also present in infected cells is probably an aggregate. Incorporation of (3)H-amino acids into each viral protein could be detected by the 2nd hr after infection and even earlier for protein N which is synthesized in the greatest amount. There was no evidence of regulation of viral protein synthesis at the transcriptive level; nonstructural and structural proteins were synthesized throughout the cycle of infection. Short pulses of (3)H-amino acids revealed no uncleaved precursor peptides that could be chased into structural peptides. Proteins N and S were chased into released virions but protein G was apparently incorporated into virions as it was being synthesized. VS viral proteins of infected cells were released by mechanically disrupting cytoplasmic membrane by nitrogen decompression and fractionated by high-speed centrifugation. Protein NS1 was present in the nonsedimentable cytoplasmic fraction throughout the cycle of infection. The nucleoprotein N was recovered primarily from the nonsedimentable fraction early in infection but aggregated into a sedimentable component, presumably the nucleocapsid, later in infection. Proteins G and S were always present in the sedimentable fraction of mechanically disrupted infected cells, presumably in association with plasma membrane. Exposure of infected cells to the membrane-dissolving agent, digitonin, resulted in solubilization of most of protein G and all of protein S but not of protein N. These experiments are compatible with the hypothesis that VS viral proteins G and S are synthesized at and inserted into plasma membrane which envelopes a nucleocapsid core to form the VS virion.
D000179:Acrylates; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001120:Arginine / Q000378:metabolism; D002247:Carbon Isotopes; D002462:Cell Membrane / Q000378:metabolism; D002642:Chick Embryo; D003593:Cytoplasm / Q000378:metabolism; D004071:Digitalis Glycosides; D004586:Electrophoresis; D005347:Fibroblasts; D005782:Gels; D005827:Genetics, Microbial; D005944:Glucosamine / Q000378:metabolism; D006023:Glycoproteins / Q000096:biosynthesis; D007739:L Cells; D007930:Leucine / Q000378:metabolism; D009698:Nucleoproteins / Q000096:biosynthesis; D013053:Spectrophotometry; D014316:Tritium; D014443:Tyrosine / Q000378:metabolism; D014721:Vesicular stomatitis Indiana virus / Q000378:metabolism*; D014764:Viral Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
D000179:Acrylates; D000596:Amino Acids; D002247:Carbon Isotopes; D004071:Digitalis Glycosides; D005782:Gels; D006023:Glycoproteins; D009698:Nucleoproteins; D014764:Viral Proteins; D014316:Tritium; D014443:Tyrosine; D001120:Arginine; D007930:Leucine; D005944:Glucosamine