title: Crystal structures of murine norovirus-1 RNA-dependent RNA polymerase.
abstract: Norovirus is one of the leading agents of gastroenteritis and is a major public health concern. In this study, the crystal structures of recombinant RNA-dependent RNA polymerase (RdRp) from murine norovirus-1 (MNV-1) and its complex with 5-fluorouracil (5FU) were determined at 2.5 Å resolution. Crystals with C2 symmetry revealed a dimer with half a dimer in the asymmetrical unit, and the protein exists predominantly as a monomer in solution, in equilibrium with a smaller population of dimers, trimers and hexamers. MNV-1 RdRp exhibited polymerization activity with a right-hand fold typical of polynucleotide polymerases. The metal ion modelled in close proximity to the active site was found to be coordinated tetrahedrally to the carboxyl groups of aspartate clusters. The orientation of 5FU observed in three molecules in the asymmetrical unit was found to be slightly different, but it was stabilized by a network of favourable interactions with the conserved active-site residues Arg185, Asp245, Asp346, Asp347 and Arg395. The information gained on the structural and functional features of MNV-1 RdRp will be helpful in understanding replication of norovirus and in designing novel therapeutic agents against this important pathogen.
authors.affiliation: Department of Biotechnology & Bioinformatics, Korea University, Chungnam 339-700, Republic of Korea.
authors.forename: Ji-Hye
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Department of Biotechnology & Bioinformatics, Korea University, Chungnam 339-700, Republic of Korea.
authors.forename: Intekhab
authors.initials: I
authors.lastname: Alam
authors.affiliation: Department of Life Science, Sogang University, Seoul 121-742, Republic of Korea.
authors.forename: Kang Rok
authors.initials: KR
authors.lastname: Han
authors.affiliation: Department of Life Science, Sogang University, Seoul 121-742, Republic of Korea.
authors.forename: Sunyoung
authors.initials: S
authors.lastname: Cho
authors.affiliation: Department of Life Science, Sogang University, Seoul 121-742, Republic of Korea.
authors.forename: Sungho
authors.initials: S
authors.lastname: Shin
authors.affiliation: Department of Biotechnology & Bioinformatics, Korea University, Chungnam 339-700, Republic of Korea.
authors.forename: Seokha
authors.initials: S
authors.lastname: Kang
authors.affiliation: Department of Life Science, Sogang University, Seoul 121-742, Republic of Korea.
authors.forename: Jai Myung
authors.initials: JM
authors.lastname: Yang
authors.affiliation: Department of Biotechnology & Bioinformatics, Korea University, Chungnam 339-700, Republic of Korea.
authors.forename: Kyung Hyun
authors.initials: KH
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D005472:Fluorouracil / Q000378:metabolism; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D029322:Norovirus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D058105:Polymerization; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D014764:Viral Proteins; D012324:RNA-Dependent RNA Polymerase; D005472:Fluorouracil