Requirement of cellular DDX3 for hepatitis C virus replication is unrelated to its interaction with the viral core protein

Angus, A.G.N. et al. (2010) Requirement of cellular DDX3 for hepatitis C virus replication is unrelated to its interaction with the viral core protein. Journal of General Virology, 91(1), pp. 122-132. (doi:10.1099/vir.0.015909-0) (PMID:19793905) (PMCID:PMC2885062)

Full text not currently available from Enlighten.

Publisher's URL: http://dx.doi.org/10.1099/vir.0.015909-0

Abstract

The cellular DEAD-box protein DDX3 was recently shown to be essential for hepatitis C virus (HCV) replication. Prior to that, we had reported that HCV core binds to DDX3 in yeast-two hybrid and transient transfection assays. Here, we confirm by co-immunoprecipitation that this interaction occurs in cells replicating the JFH1 virus. Consistent with this result, immunofluorescence staining of infected cells revealed a dramatic redistribution of cytoplasmic DDX3 by core protein to the virus assembly sites around lipid droplets. Given this close association of DDX3 with core and lipid droplets, and its involvement in virus replication, we investigated the importance of this host factor in the virus life cycle. Mutagenesis studies located a single amino acid in the N-terminal domain of JFH1 core that when changed to alanine significantly abrogated this interaction. Surprisingly, this mutation did not alter infectious virus production and RNA replication, indicating that the core–DDX3 interaction is dispensable in the HCV life cycle. Consistent with previous studies, siRNA-led knockdown of DDX3 lowered virus production and RNA replication levels of both WT JFH1 and the mutant virus unable to bind DDX3. Thus, our study shows for the first time that the requirement of DDX3 for HCV replication is unrelated to its interaction with the viral core protein.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Adair, Dr Richard and Angus, Dr Allan and Owsianka, Dr Anna and Dalrymple, Mr David and Targett-Adams, Dr Paul and Patel, Professor Arvind and McLauchlan, Professor John
Authors: Angus, A.G.N., Dalrymple, D., Boulant, S., McGivern, D.R., Clayton, R.F., Scott, M.J., Adair, R., Graham, S., Owsianka, A.M., Targett-Adams, P., Li, K., Wakita, T., McLauchlan, J., Lemon, S.M., and Patel, A.H.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Journal of General Virology
Journal Abbr.:J. Gen. Virol.
Publisher:Society for General Microbiology
ISSN:0022-1317
ISSN (Online):1465-2099
Published Online:30 September 2009

University Staff: Request a correction | Enlighten Editors: Update this record

Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
656341Virus-host interactions in hepatitis C virus infectionJohn MclauchlanMedical Research Council (MRC)MC_UU_12014/1MVLS III - CENTRE FOR VIRUS RESEARCH