Cellular casein kinase II-mediated phosphorylation of rinderpest virus P protein is a prerequisite for its role in replication/transcription of the genome.
Phosphoprotein P of rinderpest virus (RPV), when expressed in E. coli, is present in the unphosphorylated form. Bacterially expressed P protein was phosphorylated by a eukaryotic cellular extract, and casein kinase II (CK II) was identified as the cellular kinase involved in phosphorylation. In vitro phosphorylation of P-deletion mutants identified the N terminus as a phosphorylation domain. In vivo phosphorylation of single or multiple serine mutants of P protein identified serine residues at 49, 88 and 151 as phospho-acceptor residues. The role of P protein phosphorylation in virus replication/transcription was evaluated using the RPV minigenome system and replication/transcription of a reporter gene in vivo. P protein phosphorylation was shown to be essential for in vivo replication/transcription since phosphorylation-null mutants do not support expression of a reporter gene. Transfection of increased amounts of phosphorylation-null mutant did not support minigenome replication/transcription in vivo.
Kaushik
Rajnish
R
Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560012, India.
Shaila
M S
MS
Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560012, India.
D000595:Amino Acid Sequence; D000818:Animals; D047390:Casein Kinase II; D002522:Chlorocebus aethiops; D004261:DNA Replication / Q000235:genetics; D017930:Genes, Reporter; D016679:Genome, Viral; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism*; D012302:Rinderpest virus / Q000235:genetics* / Q000378:metabolism; D013379:Substrate Specificity; D014158:Transcription, Genetic / Q000235:genetics; D014709:Vero Cells; D014764:Viral Proteins / Q000378:metabolism*
D014764:Viral Proteins; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D012324:RNA-Dependent RNA Polymerase