Roles for endocytic trafficking and phosphatidylinositol 4-kinase III alpha in hepatitis C virus replication
- PMID: 19376974
- PMCID: PMC2678598
- DOI: 10.1073/pnas.0902693106
Roles for endocytic trafficking and phosphatidylinositol 4-kinase III alpha in hepatitis C virus replication
Abstract
Hepatitis C virus (HCV) reorganizes cellular membranes to establish sites of replication. The required host pathways and the mechanism of cellular membrane reorganization are poorly characterized. Therefore, we interrogated a customized small interfering RNA (siRNA) library that targets 140 host membrane-trafficking genes to identify genes required for both HCV subgenomic replication and infectious virus production. We identified 7 host cofactors of viral replication, including Cdc42 and Rock2 (actin polymerization), EEA1 and Rab5A (early endosomes), Rab7L1, and PI3-kinase C2gamma and PI4-kinase IIIalpha (phospholipid metabolism). Studies of drug inhibitors indicate actin polymerization and phospholipid kinase activity are required for HCV replication. We found extensive co-localization of the HCV replicase markers NS5A and double-stranded RNA with Rab5A and partial co-localization with Rab7L1. PI4K-IIIalpha co-localized with NS5A and double-stranded RNA in addition to being present in detergent-resistant membranes containing NS5A. In a comparison of type II and type III PI4-kinases, PI4Ks were not required for HCV entry, and only PI4K-IIIalpha was required for HCV replication. Although PI4K-IIIalpha siRNAs decreased HCV replication and virus production by almost 100%, they had no effect on initial HCV RNA translation, suggesting that PI4K-IIIalpha functions at a posttranslational stage. Electron microscopy identified the presence of membranous webs, which are thought to be the site of HCV replication, in HCV-infected cells. Pretreatment with PI4K-IIIalpha siRNAs greatly reduced the accumulation of these membranous web structures in HCV-infected cells. We propose that PI4K-IIIalpha plays an essential role in membrane alterations leading to the formation of HCV replication complexes.
Conflict of interest statement
The authors declare no conflict of interest.
Figures
Similar articles
-
Mechanisms of Cellular Membrane Reorganization to Support Hepatitis C Virus Replication.Viruses. 2016 May 20;8(5):142. doi: 10.3390/v8050142. Viruses. 2016. PMID: 27213428 Free PMC article. Review.
-
Daclatasvir inhibits hepatitis C virus NS5A motility and hyper-accumulation of phosphoinositides.Virology. 2015 Feb;476:168-179. doi: 10.1016/j.virol.2014.12.018. Epub 2014 Dec 26. Virology. 2015. PMID: 25546252 Free PMC article.
-
Daclatasvir-like inhibitors of NS5A block early biogenesis of hepatitis C virus-induced membranous replication factories, independent of RNA replication.Gastroenterology. 2014 Nov;147(5):1094-105.e25. doi: 10.1053/j.gastro.2014.07.019. Epub 2014 Jul 18. Gastroenterology. 2014. PMID: 25046163
-
Roles of phosphoinositides and phosphoinositides kinases in hepatitis C virus RNA replication.Arch Pharm Res. 2012 Oct;35(10):1701-11. doi: 10.1007/s12272-012-1001-2. Epub 2012 Nov 9. Arch Pharm Res. 2012. PMID: 23139120 Review.
-
Potential roles for cellular cofactors in hepatitis C virus replication complex formation.Commun Integr Biol. 2009 Nov;2(6):471-3. doi: 10.4161/cib.2.6.9261. Commun Integr Biol. 2009. PMID: 20195453 Free PMC article.
Cited by
-
Molecular basis for plasma membrane recruitment of PI4KA by EFR3.bioRxiv [Preprint]. 2024 Apr 30:2024.04.30.587787. doi: 10.1101/2024.04.30.587787. bioRxiv. 2024. PMID: 38746453 Free PMC article. Preprint.
-
NS5A domain I antagonises PKR to facilitate the assembly of infectious hepatitis C virus particles.PLoS Pathog. 2023 Feb 16;19(2):e1010812. doi: 10.1371/journal.ppat.1010812. eCollection 2023 Feb. PLoS Pathog. 2023. PMID: 36795772 Free PMC article.
-
Essential Domains of Oxysterol-Binding Protein Required for Poliovirus Replication.Viruses. 2022 Nov 29;14(12):2672. doi: 10.3390/v14122672. Viruses. 2022. PMID: 36560676 Free PMC article.
-
Beyond PI3Ks: targeting phosphoinositide kinases in disease.Nat Rev Drug Discov. 2023 May;22(5):357-386. doi: 10.1038/s41573-022-00582-5. Epub 2022 Nov 14. Nat Rev Drug Discov. 2023. PMID: 36376561 Free PMC article. Review.
-
Life cycle process dependencies of positive-sense RNA viruses suggest strategies for inhibiting productive cellular infection.J R Soc Interface. 2021 Nov;18(184):20210401. doi: 10.1098/rsif.2021.0401. Epub 2021 Nov 10. J R Soc Interface. 2021. PMID: 34753308 Free PMC article.
References
-
- Miyanari Y, et al. The lipid droplet is an important organelle for hepatitis C virus production. Nat Cell Biol. 2007;9:1089–1097. - PubMed
-
- Miyanari Y, et al. Hepatitis C virus non-structural proteins in the probable membranous compartment function in viral genome replication. J Biol Chem. 2003;278:50301–50308. - PubMed
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical
Molecular Biology Databases
Miscellaneous