title: Structure of hepatitis C virus polymerase in complex with primer-template RNA.
abstract: The replication of the hepatitis C viral (HCV) genome is accomplished by the NS5B RNA-dependent RNA polymerase (RdRp), for which mechanistic understanding and structure-guided drug design efforts have been hampered by its propensity to crystallize in a closed, polymerization-incompetent state. The removal of an autoinhibitory β-hairpin loop from genotype 2a HCV NS5B increases de novo RNA synthesis by >100-fold, promotes RNA binding, and facilitated the determination of the first crystallographic structures of HCV polymerase in complex with RNA primer-template pairs. These crystal structures demonstrate the structural realignment required for primer-template recognition and elongation, provide new insights into HCV RNA synthesis at the molecular level, and may prove useful in the structure-based design of novel antiviral compounds. Additionally, our approach for obtaining the RNA primer-template-bound structure of HCV polymerase may be generally applicable to solving RNA-bound complexes for other viral RdRps that contain similar regulatory β-hairpin loops, including bovine viral diarrhea virus, dengue virus, and West Nile virus.
authors.affiliation: Pharmasset Inc, Princeton, New Jersey, USA.
authors.forename: Ralph T
authors.initials: RT
authors.lastname: Mosley
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Edwards
authors.forename: Eisuke
authors.initials: E
authors.lastname: Murakami
authors.forename: Angela M
authors.initials: AM
authors.lastname: Lam
authors.forename: Rena L
authors.initials: RL
authors.lastname: Grice
authors.forename: Jinfa
authors.initials: J
authors.lastname: Du
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Sofia
authors.forename: Philip A
authors.initials: PA
authors.lastname: Furman
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Otto
mesh_terms: D002460:Cell Line; D003460:Crystallization; D004261:DNA Replication; D016174:Hepacivirus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics*; D006526:Hepatitis C / Q000821:virology; D006801:Humans; D008958:Models, Molecular; D017433:Protein Structure, Secondary; D012313:RNA / Q000235:genetics*; D013698:Templates, Genetic; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase
chemical_list: D012313:RNA; D017361:Viral Nonstructural Proteins; C034477:RNA primers; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase