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: Ralph T Mosley, Thomas E Edwards, Eisuke Murakami, Angela M Lam, Rena L Grice, Jinfa Du, Michael J Sofia, Philip A Furman, Michael J Otto
• Journal: Journal of virology (J Virol) Vol. 86 Issue 12 Pg. 6503-11 (Jun 2012) ISSN: 1098-5514 [Electronic] United States
• PMID: 22496223 (Publication Type: Journal Article)

Chemical References

• RNA primers
• Viral Nonstructural Proteins
• RNA
• NS-5 protein, hepatitis C virus

Topics

• Cell Line
• Crystallization
• DNA Replication
• Hepacivirus (chemistry, enzymology, genetics)
• Hepatitis C (virology)
• Humans
• Models, Molecular
• Protein Structure, Secondary
• RNA (genetics)
• Templates, Genetic
• Viral Nonstructural Proteins (chemistry, genetics, metabolism)