The eukaryotic translation initiation factor 3 subunit L protein interacts with FlavivirusNS5 and may modulate yellow fever virus replication
Open Access
- 22 June 2013
- journal article
- research article
- Published by Springer Science and Business Media LLC in Virology Journal
- Vol. 10 (1), 205-13
- https://doi.org/10.1186/1743-422x-10-205
Abstract
Background: Yellow fever virus (YFV) belongs to the Flavivirus genus and causes an important disease. An alarming resurgence of viral circulation and the expansion of YFV-endemic zones have been detected in Africa and South America in recent years. NS5 is a viral protein that contains methyltransferase and RNA-dependent RNA polymerase (RdRp) domains, which are essential for viral replication, and the interactions between NS5 and cellular proteins have been studied to better understand viral replication. The aim of this study was to characterize the interaction of the NS5 protein with eukaryotic translation initiation factor 3 subunit L (eIF3L) and to evaluate the role of eIF3L in yellow fever replication. Methods: To identify interactions of YFV NS5 with cellular proteins, we performed a two-hybrid screen using the YFV NS5 RdRp domain as bait with a human cDNA library, and RNApol deletion mutants were generated and analyzed using the two-hybrid system for mapping the interactions. The RNApol region involved was segmented into three fragments and analyzed using an eIF3L-expressing yeast strain. To map the NS5 residues that are critical for the interactions, we performed site-direct mutagenesis in segment 3 of the interaction domain (ID) and confirmed the interaction using in vitro assays and in vivo coimmunoprecipitation. The significance of eIF3L for YFV replication was investigated using eIF3L overexpression and RNA interference. Results: In this work, we describe and characterize the interaction of NS5 with the translation factor eIF3L. The interaction between NS5 and eIF3L was confirmed using in vitro binding and in vivo coimmunoprecipitation assays. This interaction occurs at a region (the interaction domain of the RNApol domain) that is conserved in several flaviviruses and that is, therefore, likely to be relevant to the genus. eIF3L overexpression and plaque reduction assays showed a slight effect on YFV replication, indicating that the interaction of eIF3L with YFV NS5 may play a role in YFV replication. Conclusions: Although the precise function of eIF3L on interactions with viral proteins is not entirely understood, these results indicate an interaction of eIF3L with YF NS5 and that eIF3L overexpression facilitates translation, which has potential implications for virus replication.Keywords
This publication has 32 references indexed in Scilit:
- A diverse range of gene products are effectors of the type I interferon antiviral responseNature, 2011
- The small nuclear ribonucleoprotein U1A interacts with NS5 from yellow fever virusArchiv für die gesamte Virusforschung, 2011
- Distinct Regions of Human eIF3 Are Sufficient for Binding to the HCV IRES and the 40S Ribosomal SubunitJournal of Molecular Biology, 2010
- Conserved functional domains and a novel tertiary interaction near the pseudoknot drive translational activity of hepatitis C virus and hepatitis C virus-like internal ribosome entry sitesNucleic Acids Research, 2009
- Inhibition of HIV-1 replication by eIF3fProceedings of the National Academy of Sciences of the United States of America, 2009
- Phosphorylation of yellow fever virus NS5 alters methyltransferase activityVirology, 2008
- Pathogenic flavivirusesThe Lancet, 2008
- Distinct gene expression profiles in peripheral blood mononuclear cells from patients infected with vaccinia virus, yellow fever 17D virus, or upper respiratory infectionsVaccine, 2007
- Reconstitution reveals the functional core of mammalian eIF3The EMBO Journal, 2007
- Combining genetic and biochemical approaches to identify functional molecular contact pointsBiological Procedures Online, 2006