title: Involvement of the N-terminal portion of influenza virus RNA polymerase subunit PB1 in nucleotide recognition.
abstract: The influenza virus PB1 protein functions as a catalytic subunit of the viral RNA-dependent RNA polymerase and contains the highly conserved motifs of RNA-dependent RNA polymerases together with putative nucleotide-binding sites. PB1 also binds to viral genomic RNAs and its replicative intermediates through the promoter regions. The detail function and interplay between functional domains are not clarified although a part of structures and functions of PB1 have been clarified. In this study, we analyzed the function of PB1 subunit in the sense of nucleotide recognition using ribavirin, which is a nucleoside analog and inhibits viral RNA synthesis of many RNA viruses including influenza virus. We screened ribavirin-resistant PB1 mutants from randomly mutated PB1 cDNA library using a mini-replicon assay, and we identified a single mutation at the amino acid position 27 of PB1 as an important residue for the nucleotide recognition.
authors.affiliation: Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan.
authors.forename: Nguyen Trong
authors.initials: NT
authors.lastname: Binh
authors.affiliation: Department of Infection Biology, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan.
authors.forename: Chitose
authors.initials: C
authors.lastname: Wakai
authors.affiliation: Department of Infection Biology, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan; Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kawaguchi
authors.affiliation: University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan. Electronic address: knagata@md.tsukuba.ac.jp.
authors.forename: Kyosuke
authors.initials: K
authors.lastname: Nagata
mesh_terms: D000818:Animals; D001483:Base Sequence; D004252:DNA Mutational Analysis; D004285:Dogs; D024882:Drug Resistance, Viral / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D061985:Madin Darby Canine Kidney Cells; D008727:Methotrexate / Q000494:pharmacology; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D009711:Nucleotides / Q000378:metabolism*; D021122:Protein Subunits / Q000737:chemistry* / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry* / Q000378:metabolism*; D012093:Replicon / Q000187:drug effects; D012254:Ribavirin / Q000494:pharmacology; D013329:Structure-Activity Relationship; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D050505:Mutant Proteins; D009711:Nucleotides; D021122:Protein Subunits; D014764:Viral Proteins; C077346:influenza virus polymerase basic protein 1; D012254:Ribavirin; D012324:RNA-Dependent RNA Polymerase; D008727:Methotrexate
keywords: Influenza; Nucleotide recognition; Resistant mutant; Ribavirin