The Open Biochemistry Journal




ISSN: 1874-091X ― Volume 13, 2019

The Hepatitis C Virus NS5A Stimulates NS5B During In Vitro RNA Synthesis in a Template Specific Manner



Elizabeth M Quezada, Caroline M Kane*

Abstract

The hepatitis C virus (HCV) NS5B protein contains the RNA dependent RNA polymerase (RdRp) activity that catalyzes the synthesis of the viral genome with other host and viral factors. NS5A is an HCV-encoded protein previously shown to localize to the replisome and be necessary for viral replication. However, its role in replication has not been defined. Using an in vitro biochemical assay, we detected a stimulatory effect of NS5A on the NS5B replication reaction with minimal natural templates. NS5A stimulates replication by NS5B on two templates derived from the 3’ end of the RNA genome (4 fold ± 1.3 fold). A pre-incubation step with the two proteins prior to the replication reaction and substoichiometric levels of NS5A are required for detecting stimulation. With a template derived from the 3’end complementary to the RNA genome (the negative strand) no stimulation was observed. Furthermore, with a synthetic template that allows studying different phases of replication, NS5A stimulates NS5B during elongation. These findings suggest that NS5A stimulates NS5B during synthesis of the complementary (i.e., negative) strand of the RNA genome.



Article Information


Identifiers and Pagination:

Year: 2009
Volume: 3
First Page: 39
Last Page: 48
Publisher Id: TOBIOCJ-3-39
DOI: 10.2174/1874091X00903010039

Article History:

Received Date: 11/12/2008
Revision Received Date: 11/3/2009
Acceptance Date: 12/3/2009
Electronic publication date: 20/4/2009
Collection year: 2009

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© Quezada and Kane; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.


* Address correspondence to this author at the Department of Molecular and Cell Biology, 408 Barker Hall, University of California, Berkeley, CA 94720-3202, USA; Tel: 510-642-4118; Fax: 510-642-5227; E-mail: kanecm@berkeley.edu


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