Threonine 157 of influenza virus PA polymerase subunit modulates RNA replication in infectious viruses.
Previous results have shown a correlation between the decrease in protease activity of several influenza A virus PA protein mutants and the capacity to replicate of the corresponding mutant ribonucleoproteins (RNPs) reconstituted in vivo. In this work we studied the phenotype of mutant viruses containing these mutations. Viruses with a T162A mutation, which showed a very moderate decrease both in protease and replication activities of reconstituted RNPs, showed a wild-type phenotype. Viruses with a T157A mutation, which presented a severe decrease in protease activity and replication of RNPs, showed a complex phenotype: (i) transport to the nucleus of PAT157A protein was delayed, (ii) virus multiplication was reduced at both low and high multiplicities, (iii) transcriptive synthesis was unaltered while replicative synthesis, especially cRNA, was diminished, and (iv) viral pathogenesis in mice was reduced, as measured by loss of body weight and virus titers in lungs. Finally, recombinant viruses with a T157E mutation in PA protein, which resulted in a drastic reduction of protease and replication activities of RNPs, were not viable. These results indicate that residue T157 in PA protein is important for the capacity of viral polymerase to synthesize cRNA.
Centro Nacional de Biotecnología, Cantoblanco, 28049 Madrid, Spain.
Maite
M
Huarte
Ana
A
Falcón
Yuri
Y
Nakaya
Juan
J
Ortín
Adolfo
A
García-Sastre
Amelia
A
Nieto
D000818:Animals; D002460:Cell Line; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004285:Dogs; D015967:Gene Expression Regulation, Viral; D006801:Humans; D009980:Influenza A virus / Q000201:enzymology / Q000472:pathogenicity* / Q000502:physiology; D007251:Influenza, Human / Q000503:physiopathology / Q000821:virology; D051379:Mice; D009154:Mutation; D012367:RNA, Viral / Q000096:biosynthesis*; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism; D013912:Threonine; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication
C061290:PA protein, influenza viruses; D012367:RNA, Viral; D012261:Ribonucleoproteins; D014764:Viral Proteins; D013912:Threonine; D012324:RNA-Dependent RNA Polymerase; D012321:DNA-Directed RNA Polymerases