title: Specificity and functional interaction of the polymerase complex proteins of human and avian metapneumoviruses.
abstract: Human metapneumovirus (HMPV) and avian metapneumovirus (AMPV) have a similar genome organization and protein composition, but a different host range. AMPV subgroup C (AMPV-C) is more closely related to HMPV than other AMPVs. To investigate the specificity and functional interaction of the polymerase complex proteins of human and avian metapneumoviruses, a minireplicon system was generated for AMPV-C and used in combination with minireplicon systems for HMPV lineages A1 and B1. Viral RNA-like molecules representing HMPV-A1 and -B1, AMPV-A and -C and human respiratory syncytial virus were replicated efficiently by polymerase complexes of HMPV-A1 and -B1 and AMPV-C, but not by polymerase complexes of bovine parainfluenza virus 3. Upon exchange of HMPV and AMPV-C polymerase complex components, all chimeric polymerase complexes were functional; exchange between HMPVs did not result in altered polymerase activity, whereas exchange between HMPVs and AMPV-C did. Recombinant HMPV-B1 viruses in which polymerase genes were exchanged with those of HMPV-A1 replicated with normal kinetics in vitro, whilst replacement with AMPV-C genes resulted in moderate differences in virus replication. In hamsters, recombinant HMPV-B1 viruses in which individual polymerase genes were exchanged with those of AMPV-C were attenuated, irrespective of the results obtained with minireplicon systems or in vitro replication assays. This study provides insight into the specificity and functional interaction of polymerase complex proteins of human and avian metapneumoviruses, but neither minireplicon systems nor in vitro replication kinetics were found to be predictive for attenuation in permissive animals.
authors.affiliation: Department of Virology, Erasmus MC, PO Box 2040, 3000 CA Rotterdam, The Netherlands.
authors.forename: Miranda
authors.initials: M
authors.lastname: de Graaf
authors.affiliation: Department of Virology, Erasmus MC, PO Box 2040, 3000 CA Rotterdam, The Netherlands.
authors.forename: Sander
authors.initials: S
authors.lastname: Herfst
authors.affiliation: Department of Virology, Erasmus MC, PO Box 2040, 3000 CA Rotterdam, The Netherlands.
authors.forename: Eefje J A
authors.initials: EJA
authors.lastname: Schrauwen
authors.affiliation: Department of Virology, Erasmus MC, PO Box 2040, 3000 CA Rotterdam, The Netherlands.
authors.forename: Ying
authors.initials: Y
authors.lastname: Choi
authors.affiliation: Department of Virology, Erasmus MC, PO Box 2040, 3000 CA Rotterdam, The Netherlands.
authors.forename: Bernadette G
authors.initials: BG
authors.lastname: van den Hoogen
authors.affiliation: Department of Virology, Erasmus MC, PO Box 2040, 3000 CA Rotterdam, The Netherlands.
authors.forename: Albert D M E
authors.initials: ADME
authors.lastname: Osterhaus
authors.affiliation: Department of Virology, Erasmus MC, PO Box 2040, 3000 CA Rotterdam, The Netherlands.
authors.forename: Ron A M
authors.initials: RAM
authors.lastname: Fouchier
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D006224:Cricetinae; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006801:Humans; D008647:Mesocricetus; D029121:Metapneumovirus / Q000502:physiology*; D018184:Paramyxoviridae Infections / Q000821:virology*; D011995:Recombination, Genetic; D012093:Replicon; D013045:Species Specificity; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D014764:Viral Proteins; D012321:DNA-Directed RNA Polymerases