title: Host-range determinants on the PB2 protein of influenza A viruses control the interaction between the viral polymerase and nucleoprotein in human cells.
abstract: The transcription/replication activity of ribonucleoproteins derived from influenza A primary isolates of human (A/Paris/908/97) or avian origin (A/Mallard/Marquenterre/MZ237/83, A/Hong Kong/156/97) was compared upon reconstitution in mammalian or avian cells, using viral-like reporter RNAs synthesized under the control of the human and chicken RNA polymerase I promoters, respectively. In avian cells, transcription/replication activities were in the same range with all ribonucleoproteins tested. In human cells, ribonucleoproteins derived from A/Mallard/Marquenterre/MZ237/83 showed reduced transcription/replication activity and reduced NP binding to the PB1-PB2-PA complex (P) or to the isolated PB2 subunit, as compared to the ribonucleoproteins derived from A/Paris/908/97. Both defects were restored when PB2 residue Glu-627 was changed to a Lys. Ribonucleoproteins derived from the human A/Hong Kong/156/97 H5N1 isolate showed efficient NP-P interaction in human cells, and high levels of activity which were determined mostly by the PB2 and PA proteins. Our data suggest that PB2 might play a pivotal role in molecular interactions involving both the viral nucleoprotein and cellular proteins.
authors.affiliation: Unité de Génétique Moléculaire des Virus Respiratoires, URA 1966 CNRS, EA302 Université Paris 7, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris Cedex 15, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Labadie
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Dos Santos Afonso
authors.forename: Marie-Anne
authors.initials: MA
authors.lastname: Rameix-Welti
authors.forename: Sylvie
authors.initials: S
authors.lastname: van der Werf
authors.forename: Nadia
authors.initials: N
authors.lastname: Naffakh
mesh_terms: D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002645:Chickens; D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis / Q000235:genetics; D002522:Chlorocebus aethiops; D017930:Genes, Reporter; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000502:physiology; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics / Q000502:physiology; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000502:physiology; D009980:Influenza A virus / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D025941:Protein Interaction Mapping; D012318:RNA Polymerase I; D012367:RNA, Viral / Q000096:biosynthesis; D012261:Ribonucleoproteins / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism*; D014779:Virus Replication
chemical_list: C502893:PB2 protein, influenza virus; D012367:RNA, Viral; D012261:Ribonucleoproteins; D014764:Viral Proteins; D015500:Chloramphenicol O-Acetyltransferase; D012318:RNA Polymerase I