World J Hepatol. 2016 Jul 08;8(19):796-814. doi: 10.4254/wjh.v8.i19.796.
Assembly and release of infectious hepatitis C virus involving unusual organization of the secretory pathway.
World journal of hepatology
Miriam Triyatni, Edward A Berger, Bertrand Saunier
Affiliations
Affiliations
- Miriam Triyatni, Roche Innovation Center, Roche Pharma Research and Early Development, F. Hoffmann-La Roche Ltd., CH-4070 Basel, Switzerland.
PMID: 27429716
PMCID: PMC4937168 DOI: 10.4254/wjh.v8.i19.796
Abstract
AIM: To determine if calnexin (CANX), RAB1 and alpha-tubulin were involved in the production of hepatitis C virus (HCV) particles by baby hamster kidney-West Nile virus (BHK-WNV) cells.
METHODS: Using a siRNA-based approach complemented with immuno-fluorescence confocal microscope and Western blot studies, we examined the roles of CANX, RAB1 and alpha-tubulin in the production of HCV particles by permissive BHK-WNV cells expressing HCV structural proteins or the full-length genome of HCV genotype 1a. Immuno-fluorescence studies in producer cells were performed with monoclonal antibodies against HCV structural proteins, as well as immunoglobulin from the serum of a patient recently cured from an HCV infection of same genotype. The cellular compartment stained by the serum immunoglobulin was also observed in thin section transmission electron microscopy. These findings were compared with the JFH-1 strain/Huh-7.5 cell model.
RESULTS: We found that CANX was necessary for the production of HCV particles by BHK-WNV cells. This process involved the recruitment of a subset of HCV proteins, detected by immunoglobulin of an HCV-cured patient, in a compartment of rearranged membranes bypassing the endoplasmic reticulum-Golgi intermediary compartment and surrounded by mitochondria. It also involved the maturation of N-linked glycans on HCV envelope proteins, which was required for assembly and/or secretion of HCV particles. The formation of this specialized compartment required RAB1; upon expression of HCV structural genes, this compartment developed large vesicles with viral particles. RAB1 and alpha-tubulin were required for the release of HCV particles. These cellular factors were also involved in the production of HCVcc in the JFH-1 strain/Huh-7.5 cell system, which involves HCV RNA replication. The secretion of HCV particles by BHK-WNV cells presents similarities with a pathway involving caspase-1; a caspase-1 inhibitor was found to suppress the production of HCV particles from a full-length genome.
CONCLUSION: Prior activity of the WNV subgenomic replicon in BHK-21 cells promoted re-wiring of host factors for the assembly and release of infectious HCV in a caspase-1-dependent mechanism.
Keywords: Flavivirus replicon; Hepatitis C virus; Host cellular factors; Membrane rearrangements; Virus assembly and secretion
References
- J Virol. 2012 Jun;86(12):6491-502 - PubMed
- Hepatology. 2011 Oct;54(4):1149-56 - PubMed
- J Virol. 1996 Feb;70(2):778-86 - PubMed
- J Virol. 2002 Sep;76(18):9335-44 - PubMed
- Eur J Immunol. 2004 Oct;34(10):2834-42 - PubMed
- J Virol. 2015 Mar;89(5):2956-61 - PubMed
- PLoS Pathog. 2014 Oct 02;10(10):e1004424 - PubMed
- PLoS One. 2014 Dec 01;9(12):e113034 - PubMed
- Cell Microbiol. 2013 Jan;15(1):24-34 - PubMed
- Trends Microbiol. 2011 Feb;19(2):95-103 - PubMed
- Science. 2015 Jul 10;349(6244):195-8 - PubMed
- Annu Rev Biochem. 2004;73:1019-49 - PubMed
- Nat Rev Microbiol. 2013 Oct;11(10):688-700 - PubMed
- Biochim Biophys Acta. 2014 Apr;1837(4):461-9 - PubMed
- EMBO J. 2009 Apr 22;28(8):1016-28 - PubMed
- J Virol. 2010 Nov;84(21):10999-1009 - PubMed
- Cytokine. 2015 Aug;74(2):213-8 - PubMed
- Science. 1997 Jul 25;277(5325):570-4 - PubMed
- J Virol. 2008 Aug;82(15):7624-39 - PubMed
- Virology. 2006 Mar 1;346(1):53-65 - PubMed
- Science. 1998 Jul 10;281(5374):269-72 - PubMed
- PLoS Pathog. 2009 Mar;5(3):e1000339 - PubMed
- Science. 2000 Jul 21;289(5478):444-8 - PubMed
- J Virol. 2014 Feb;88(3):1433-46 - PubMed
- Traffic. 2010 May;11(5):616-25 - PubMed
- Hepatology. 2015 Nov;62(5):1346-52 - PubMed
- Viruses. 2015 Jun 19;7(6):3204-25 - PubMed
- Cell Logist. 2014 Jan 09;3:e27687 - PubMed
- J Cell Sci. 2010 Jul 1;123(Pt 13):2273-80 - PubMed
- J Cell Biol. 2011 Nov 14;195(4):605-15 - PubMed
- J Biol Chem. 2005 Jun 17;280(24):23349-55 - PubMed
- Science. 1999 Jul 2;285(5424):110-3 - PubMed
- J Virol. 2010 Jan;84(2):773-87 - PubMed
- PLoS Pathog. 2009 Jun;5(6):e1000475 - PubMed
- Sci Rep. 2014 Dec 08;4:7357 - PubMed
- Cell. 2009 Aug 7;138(3):549-61 - PubMed
- Nat Rev Microbiol. 2008 Sep;6(9):699-708 - PubMed
- Lancet Infect Dis. 2005 Sep;5(9):558-67 - PubMed
- J Virol. 2006 Mar;80(6):2832-41 - PubMed
- Proc Natl Acad Sci U S A. 2009 May 5;106(18):7577-82 - PubMed
- Cell Host Microbe. 2009 Apr 23;5(4):365-75 - PubMed
- World J Gastroenterol. 2016 Feb 14;22(6):1953-65 - PubMed
- J Virol. 2007 Aug;81(16):8374-83 - PubMed
- Mol Biol Cell. 2006 Aug;17(8):3494-507 - PubMed
- J Gen Virol. 2001 Aug;82(Pt 8):1877-83 - PubMed
- Proc Natl Acad Sci U S A. 1980 Feb;77(2):780-4 - PubMed
- PLoS Pathog. 2015 Sep 25;11(9):e1005185 - PubMed
- Lancet. 2015 Mar 21;385(9973):1124-35 - PubMed
- Nature. 2009 Aug 20;460(7258):978-83 - PubMed
- Nat Rev Mol Cell Biol. 2009 Feb;10(2):148-55 - PubMed
- Cell Mol Life Sci. 2002 Jan;59(1):112-25 - PubMed
- PLoS Pathog. 2009 Oct;5(10):e1000632 - PubMed
- Nat Med. 2005 May;11(5):522-30 - PubMed
- Proc Natl Acad Sci U S A. 2011 Aug 30;108(35):14590-5 - PubMed
- Proc Natl Acad Sci U S A. 2006 Mar 7;103(10):3805-9 - PubMed
- Gastroenterology. 2010 Nov;139(5):1774-83, 1783.e1-6 - PubMed
- Hepatology. 2007 Aug;46(2):330-8 - PubMed
- PLoS Pathog. 2011 Jul;7(7):e1002144 - PubMed
- Nat Cell Biol. 2007 Sep;9(9):1089-97 - PubMed
- Curr Opin Microbiol. 2014 Aug;20:1-9 - PubMed
- J Virol. 2004 Jul;78(14):7737-47 - PubMed
- J Clin Invest. 2010 Mar;120(3):650-3 - PubMed
- J Viral Hepat. 2013 Jun;20(6):369-76 - PubMed
- J Hepatol. 2014 Nov;61(1 Suppl):S34-44 - PubMed
- J Virol. 2008 Nov;82(21):10519-31 - PubMed
- PLoS Pathog. 2007 Aug 31;3(8):e108 - PubMed
- Hepatology. 2012 Sep;56(3):861-72 - PubMed
- Sci Rep. 2015 Jul 10;5:10592 - PubMed
- PLoS Pathog. 2011 Apr;7(4):e1001333 - PubMed
- Mol Syst Biol. 2008;4:230 - PubMed
- Cell Host Microbe. 2007 Oct 11;2(4):229-39 - PubMed
- J Virol. 2006 Nov;80(22):11074-81 - PubMed
- BMJ. 2014 Jul 07;348:g3308 - PubMed
- Glycobiology. 2013 Apr;23(4):453-74 - PubMed
- J Virol. 2003 Jan;77(1):546-59 - PubMed
- Nat Commun. 2014 Sep 16;5:4874 - PubMed
- Cell Host Microbe. 2013 Nov 13;14(5):522-34 - PubMed
- Cold Spring Harb Perspect Biol. 2014 Oct 16;6(12 ):a016287 - PubMed
- Nat Med. 2005 Jul;11(7):791-6 - PubMed
- J Biol Chem. 2009 May 15;284(20):13778-91 - PubMed
- J Virol. 2002 Jul;76(14):6919-28 - PubMed
- J Virol. 2003 Mar;77(5):3181-90 - PubMed
- Lancet Infect Dis. 2015 Apr;15(4):451-60 - PubMed
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