Kinetics of synthesis and phosphorylation of respiratory syncytial virus polypeptides.
The kinetics of synthesis of [35S]methionine-labelled respiratory syncytial virus-specific proteins were studied in CV-1 cells infected at high multiplicity. Immunoprecipitated viral proteins resolved by SDS-PAGE were quantified by scanning fluorographs of protein bands. The nucleocapsid (N) protein was detectable by 2 h post-infection (p.i.), whereas the phospho- (P), matrix (M) and fusion (Fo) proteins and Vp24 (a matrix-like protein) were first detected between 4 and 6 h p.i. Synthesis of the glyco- (G) protein was first detected at 6 h p.i. and reached its peak synthesis rate at 10 h p.i. Virus-specific P, M and Vp24 proteins were phosphorylated in infected cells. The P protein was highly phosphorylated in purified virions whereas phosphorylated species of the M and Vp24 proteins were minor components. The phosphorylated form of the P protein was detected by monoclonal antibody precipitation, confirming the identity of this protein. The N protein was not phosphorylated in infected cells or in virions. Synthesis of [35S]methionine-labelled proteins preceded detectable 32Pi labelling by several hours. The putative phosphorylated M protein was detected at 6 h p.i. before phosphorylated forms of P and Vp24 were seen. The timing of appearance of the phosphorylated species of P and Vp24 proteins in infected cells corresponded to the release of infectious virions from infected cell monolayers at 10 to 12 h p.i.
Lambert
D M
DM
Department of Molecular Virology, James N. Gamble Institute of Medical Research, Cincinnati, Ohio 45219.
Hambor
J
J
Diebold
M
M
Galinski
B
B
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D002460:Cell Line; D002522:Chlorocebus aethiops; D007700:Kinetics; D010452:Peptide Biosynthesis; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D012136:Respiratory Syncytial Viruses / Q000276:immunology / Q000378:metabolism*; D014764:Viral Proteins / Q000096:biosynthesis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D014764:Viral Proteins