Translational control by influenza virus. Selective and cap-dependent translation of viral mRNAs in infected cells.
In cells infected by influenza virus type A, host protein synthesis undergoes a rapid and dramatic shutoff. To define the molecular mechanisms underlying this selective translation, a transfection/infection protocol was developed utilizing viral and cellular cDNA clones. When COS-1 cells were transfected with cDNAs encoding nonviral genes and subsequently infected with influenza virus, protein expression from the exogenous genes was diminished, similar to the endogenous cellular genes. However, when cells were transfected with a truncated influenza viral nucleocapsid protein (NP-S) gene, the NP-S protein was made as efficiently in influenza virus infected cells as in uninfected cells, showing that the NP-S mRNA, although expressed independently of the influenza virus replication machinery, was still recognized as a viral and not a cellular mRNA. Northern blot analysis demonstrated that the selective blocks to nonviral protein synthesis were at the level of translation. Moreover, polysome experiments revealed that the translational blocks occurred at both the initiation and elongation stages of cellular protein synthesis. Finally, we utilized this transfection/infection system as well as double infection experiments to demonstrate that the translation of influenza viral mRNAs probably occurred in a cap-dependent manner as poliovirus infection inhibited influenza viral mRNA translation.
Garfinkel
M S
MS
Department of Microbiology, School of Medicine, University of Washington, Seattle 98195.
Katze
M G
MG
D002213:Capsid / Q000235:genetics; D002460:Cell Line; D004247:DNA / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D009980:Influenza A virus / Q000235:genetics*; D014176:Protein Biosynthesis; D012315:RNA Caps; D012333:RNA, Messenger / Q000235:genetics*; D012367:RNA, Viral / Q000235:genetics; D014162:Transfection; D014758:Viral Core Proteins / Q000235:genetics
D012315:RNA Caps; D012333:RNA, Messenger; D012367:RNA, Viral; D014758:Viral Core Proteins; D004247:DNA