Hepatitis C viral NS3-4A protease activity is enhanced by the NS3 helicase.

The Journal of Biological Chemistry
Rudolf K Beran, A M Pyle

Abstract

Non-structural protein 3 (NS3) is a multifunctional enzyme possessing serine protease, NTPase, and RNA unwinding activities that are required for hepatitis C viral (HCV) replication. HCV non-structural protein 4A (NS4A) binds to the N-terminal NS3 protease domain to stimulate NS3 serine protease activity. In addition, the NS3 protease domain enhances the RNA binding, ATPase, and RNA unwinding activities of the C-terminal NS3 helicase domain (NS3hel). To determine whether NS3hel enhances the NS3 serine protease activity, we purified truncated and full-length NS3-4A complexes and examined their serine protease activities under a variety of salt and pH conditions. Our results indicate that the helicase domain enhances serine protease activity, just as the protease domain enhances helicase activity. Thus, the two enzymatic domains of NS3-4A are highly interdependent. This is the first time that such a complete interdependence has been demonstrated for a multifunctional, single chain enzyme. NS3-4A domain interdependence has important implications for function during the viral lifecycle as well as for the design of inhibitor screens that target the NS3-4A protease.

References

Mar 15, 1996·The Journal of Biological Chemistry·C SteinkühlerR De Francesco
Mar 27, 1999·Nature·S J Li, M Hochstrasser
May 14, 1999·Trends in Biochemical Sciences·J de la CruzP Linder
Aug 2, 2000·Proceedings of the National Academy of Sciences of the United States of America·S K SinghM R Maurizi
Aug 30, 2005·Nature Immunology·Taro KawaiShizuo Akira
Sep 13, 2005·Molecular Cell·Liang-Guo XuHong-Bing Shu
Dec 15, 2005·Journal of Virology·Angela M I Lam, David N Frick
Jun 19, 2007·Current Opinion in Structural Biology·Eckhard Jankowsky, Margaret E Fairman

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Citations

Oct 28, 2010·Archives of Virology·Sergey A ShiryaevAlex Y Strongin
Mar 14, 2009·Proceedings of the National Academy of Sciences of the United States of America·V SchregelN Tautz
Dec 14, 2011·Proceedings of the National Academy of Sciences of the United States of America·Nikolaus SchieringUrsula Bodendorf
Apr 7, 2010·The Journal of Biological Chemistry·Vaishnavi RajagopalSmita S Patel
May 12, 2010·The Journal of Biological Chemistry·Kevin D RaneyCraig E Cameron
Oct 13, 2010·Antimicrobial Agents and Chemotherapy·Chi-Chen YangAndrew Yueh
Jan 19, 2012·Antimicrobial Agents and Chemotherapy·Christoph WelschIris Antes
Jan 21, 2009·Journal of Virology·Rudolf K F BeranAnna Marie Pyle
May 5, 2012·PloS One·Leopoldo G GebhardAndrea V Gamarnik
Jun 30, 2009·The Tohoku Journal of Experimental Medicine·Alejandro Daniel Bermúdez-AguirreJulio Reyes-Leyva
Feb 18, 2010·Future Virology·Craig A Belon, David N Frick
Oct 2, 2012·Nature Chemical Biology·Susanne M Saalau-BethellHarren Jhoti
Apr 20, 2010·FEBS Letters·Zhi-Shun HuangHuey-Nan Wu
Apr 23, 2013·Current Opinion in Virology·Meigang Gu, Charles M Rice
Dec 11, 2014·Proteomics·Veronica M JarockiSteven P Djordjevic
Jan 5, 2011·Journal of Molecular Recognition : JMR·Matthis GeitmannU Helena Danielson
Oct 15, 2013·Protein Science : a Publication of the Protein Society·Cihan AydinCelia A Schiffer
Jun 12, 2009·Clinical Science·Hengli Tang, Henry Grisé
Jul 21, 2012·Virus Research·Robert VaughanC Cheng Kao
Sep 11, 2012·Current Opinion in Virology·Laurent Chatel-ChaixDaniel Lamarre
Mar 19, 2016·Journal of Medical Virology·Nicola CoppolaEvangelista Sagnelli
Aug 1, 2010·Viruses·Laurent Chatel-ChaixDaniel Lamarre
Aug 25, 2017·Journal of Virology·Fengwei ZhengZishu Pan
Feb 6, 2015·Journal of Virology·M Alejandra TortoriciFélix A Rey

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Methods Mentioned

BETA
PCR
gel filtration
protease assay
electrophoresis

Software Mentioned

Systat
Sigma Plot

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