NS3 from Hepatitis C Virus Strain JFH-1 Is an Unusually Robust Helicase That Is Primed To Bind and Unwind Viral RNA

Journal of Virology
Ting ZhouAnna Marie Pyle

Abstract

Hepatitis C viruses (HCV) encode a helicase enzyme that is essential for viral replication and assembly (nonstructural protein 3 [NS3]). This helicase has become the focus of extensive basic research on the general helicase mechanism, and it is also of interest as a novel drug target. Despite the importance of this protein, mechanistic work on NS3 has been conducted almost exclusively on variants from HCV genotype 1. Our understanding of NS3 from the highly active HCV strains that are used to study HCV genetics and mechanism in cell culture (such as JFH-1) is lacking. We therefore set out to determine whether NS3 from the replicatively efficient genotype 2a strain JFH-1 displays novel functional or structural properties. Using biochemical assays for RNA binding and duplex unwinding, we show that JFH-1 NS3 binds RNA much more rapidly than the previously studied NS3 variants from genotype 1b. Unlike NS3 variants from other genotypes, JFH-1 NS3 binds RNA with high affinity in a functionally active form that is capable of immediately unwinding RNA duplexes without undergoing rate-limiting conformational changes that precede activation. Unlike other superfamily 2 (SF2) helicases, JFH-1 NS3 does not require long 3' overhangs, and it ...Continue Reading

References

Jul 25, 1995·Nucleic Acids Research·F WincottN Usman
Jun 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·I Wong, T M Lohman
Jul 1, 1999·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·M R BeardS M Lemon
Nov 5, 1999·The Journal of Biological Chemistry·M K Levin, S S Patel
Aug 29, 2003·The Journal of Biological Chemistry·Angela M I LamDavid N Frick
Apr 17, 2004·The Journal of Biological Chemistry·Mikhail K LevinSmita S Patel
Jul 23, 2004·Nature·Victor Serebrov, Anna Marie Pyle
Nov 13, 2004·Nature·Martin R SingletonDale B Wigley
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·Paul Emsley, Kevin Cowtan
Jan 7, 2005·The Journal of Biological Chemistry·Alan J TackettKevin D Raney
Apr 5, 2005·Nature Structural & Molecular Biology·Mikhail K LevinSmita S Patel
Jun 11, 2005·Science·Brett D LindenbachCharles M Rice
Jun 25, 2008·Annual Review of Biophysics·Anna Marie Pyle
Aug 30, 2008·The Journal of Biological Chemistry·Rudolf K F Beran, Anna Marie Pyle
Oct 11, 2008·Journal of the American Chemical Society·Zachary R LaughreyMarcey L Waters
Nov 18, 2008·The Journal of Biological Chemistry·Victor SerebrovAnna Marie Pyle
Jan 20, 2009·Bioinformatics·Andrew M WaterhouseGeoffrey J Barton
Aug 1, 2007·Journal of Applied Crystallography·Airlie J McCoyRandy J Read
Feb 4, 2010·Acta Crystallographica. Section D, Biological Crystallography·Paul D AdamsPeter H Zwart
Apr 7, 2010·The Journal of Biological Chemistry·Vaishnavi RajagopalSmita S Patel
Dec 15, 2010·Journal of Molecular Biology·Todd C ApplebyJohn R Somoza
Apr 3, 2012·Proceedings of the National Academy of Sciences of the United States of America·Yi-Ping LiJens Bukh
Jun 21, 2012·Methods in Enzymology·Steve C Ding, Anna Marie Pyle
Oct 2, 2012·Nature Chemical Biology·Susanne M Saalau-BethellHarren Jhoti
Oct 12, 2013·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Donald B SmithPeter Simmonds
Mar 1, 2016·Molecular Cell·Nathan PirakitikulrAnna M Pyle

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Citations

Oct 2, 2019·Cold Spring Harbor Perspectives in Medicine·Keisuke TabataRalf Bartenschlager
Sep 11, 2019·Cold Spring Harbor Perspectives in Medicine·Takaji Wakita
Apr 4, 2021·Viruses·Hui-Chun LiShih-Yen Lo

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Datasets Mentioned

BETA
AF139594
BAB32872

Methods Mentioned

BETA
equilibrium filter binding
size exclusion chromatography
PCA
gel filtration
dot blot
fluorescence spectroscopy

Software Mentioned

Clustal Omega
ImageQuant
JalView
PyMOL
PHASER
MatLab
ProtParam
Rapid Automated Processing of Data ( RAPD )
PHENIX
XDS

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