Importance of the cysteine-rich carboxyl-terminal half of V protein for Sendai virus pathogenesis.
The Sendai virus V protein is a nonstructural trans-frame protein whose cysteine-rich C-terminal half is fused to the acidic N-terminal half of the P protein via mRNA editing. We recently created a mutant by disrupting the editing motif, which is devoid of mRNA editing and hence unable to produce the V protein, and demonstrated that this V(-) virus replicated normally or even faster with augmented gene expression and cytopathogenicity in cells in vitro, but was strongly attenuated in pathogenicity for mice (A. Kato, K. Kiyotani, Y. Sakai, T. Yoshida, and Y. Nagai, EMBO J. 16:578-587, 1997). Thus, although categorized as a nonessential protein, the V protein appeared to encode a luxury function required for the viral in vivo pathogenesis. Here, we created another version of a V-deficient mutant, VdeltaC, encoding only the N-terminal half but not the V-specific C-terminal half, by introducing a stop codon in the trans-V frame, and then we compared its in vitro and in vivo phenotypes with those of the V(-) and wild-type viruses. The VdeltaC virus was found to be similar to the wild-type virus in vitro with no augmented gene expression and cytopathogenicity, but in vivo, it resembled the V(-) virus, displaying a similarly attenuated phenotype. Thus, the pathogenicity determinant in the V protein was mapped to the C-terminal half. The N-terminal half was likely sufficient to confer normal (wild-type) in vitro phenotypes.
Kato
A
A
Department of Viral Infection, Institute of Medical Science, University of Tokyo, Japan.
Kiyotani
K
K
Sakai
Y
Y
Yoshida
T
T
Shioda
T
T
Nagai
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003545:Cysteine; D017931:DNA Primers; D018076:DNA, Complementary; D007668:Kidney; D008168:Lung / Q000821:virology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D016254:Mutagenesis, Insertional; D016297:Mutagenesis, Site-Directed; D017393:RNA Editing; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D018108:Respirovirus / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology*; D010253:Respirovirus Infections / Q000503:physiopathology*; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000502:physiology*; D014779:Virus Replication; D015430:Weight Gain
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; D003545:Cysteine