Role of primary constitutive phosphorylation of Sendai virus P and V proteins in viral replication and pathogenesis.
Functional analysis of the primary constitutive phosphorylation of Sendai virus P and V proteins was performed using both in vitro and in vivo systems. Sendai virus minigenome transcription and replication in transfected cells were not significantly affected in the presence of primary phosphorylation deficient P protein (S249A, S249D, P250A) as measured by either the luciferase activity or the Northern blot analysis. Similarly, recombinant Sendai viruses lacking the primary phosphorylation in P grew to titers close to the wild-type virus in cell cultures and in the natural host of Sendai virus, the mouse. Mutant viruses showed no altered pathogenesis in mice lungs. Oligomerization of P by binding WT P or mutant P to GST-P (WT) Sepharose beads revealed that the primary phosphorylation was not crucial for P protein oligomerization. Similar to P protein primary phosphorylation, the V protein primary phosphorylation at serine249 was not essential for minigenome transcription and replication, as both WT and mutant V proteins were found equally inhibitory to the minigenome replication. These results show that the primary phosphorylation of P protein has no essential role in Sendai virus transcription, replication, and pathogenesis.
Hu
C J
CJ
Department of Immunology/Microbiology, Rush-Presbyterian-St. Luke's Medical Center, Chicago, Illinois 60612, USA.
Kato
A
A
Bowman
M C
MC
Kiyotani
K
K
Yoshida
T
T
Moyer
S A
SA
Nagai
Y
Y
Gupta
K C
KC
D000818:Animals; D015152:Blotting, Northern; D002460:Cell Line; D016679:Genome, Viral; D006801:Humans; D008168:Lung / Q000821:virology; D008171:Lung Diseases / Q000821:virology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D012367:RNA, Viral / Q000096:biosynthesis; D018108:Respirovirus / Q000235:genetics / Q000502:physiology*; D010253:Respirovirus Infections / Q000473:pathology / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication
C055859:P protein, Sendai virus; D010750:Phosphoproteins; D012367:RNA, Viral; C111739:V protein, Sendai virus; D014764:Viral Proteins