Human parainfluenza virus type 2 V protein inhibits genome replication by binding to the L protein: possible role in promoting viral fitness.
The human parainfluenza virus type 2 (hPIV2) V protein plays important roles in inhibiting the host interferon response and promoting virus growth, but its role in hPIV2 replication and transcription is not clear. A green fluorescent protein (GFP)-expressing a negative-sense minigenomic construct of hPIV2 has been established by standard technology, with helper plasmids expressing the nucleocapsid protein (NP), phosphoprotein (P), and large RNA polymerase (L) protein, to examine the role of V protein. We found that the simultaneous expression of wild-type V protein in the minigenome system inhibited GFP expression, at least in part, by inhibiting minigenome replication. In contrast, expression of C terminally truncated or mutant hPIV2 V proteins had no effect. Moreover, the V protein of simian virus 41, the rubulavirus most closely related virus to hPIV2, also inhibited GFP expression, whereas that of PIV5, a more distantly related rubulavirus, did not. Using these other rubulavirus V proteins, as well as various mutant hPIV2 V proteins, we found that the ability of V protein to inhibit GFP expression correlated with its ability to bind to L protein via its C-terminal V protein-specific region, but there was no correlation with NP binding. A possible role for this inhibition of genome replication in promoting viral fitness is discussed.
Nishio
Machiko
M
Department of Microbiology, Mie University Graduate School of Medicine, 2-174, Edobashi, Tsu, Mie 514-8507, Japan. nishio@doc.medic.mie-u.ac.jp
Ohtsuka
Junpei
J
Tsurudome
Masato
M
Nosaka
Tetsuya
T
Kolakofsky
Daniel
D
D000818:Animals; D000911:Antibodies, Monoclonal; D019556:COS Cells; D002522:Chlorocebus aethiops; D017931:DNA Primers / Q000235:genetics; D004261:DNA Replication / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D047468:Immunoprecipitation; D009154:Mutation / Q000235:genetics; D010223:Parainfluenza Virus 2, Human; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020798:Two-Hybrid System Techniques; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication / Q000235:genetics*
D000911:Antibodies, Monoclonal; D017931:DNA Primers; C068854:V protein, human parainfluenza virus type 1; D014764:Viral Proteins; C069129:L protein, Human parainfluenza virus 2; D049452:Green Fluorescent Proteins