Top

Virology Journal

Published in:

Open Access 01-12-2016 | Research

Disruption of clathrin-dependent trafficking results in the failure of grass carp reovirus cellular entry

Authors: Hao Wang, Weisha Liu, Fei Yu, Liqun Lu

Published in: Virology Journal | Issue 1/2016

Abstract

Background

Grass carp reovirus (GCRV) is responsible for viral hemorrhagic disease in cultured grass carp (Ctenopharyngon idellus). GCRV is a non-enveloped, double-stranded RNA virus in the genus Aquareovirus, of the family Reoviridae, which encodes seven structural proteins (VP1-VP7) and five nonstructural proteins (NS80, NS38, NS31, NS26, and NS16). To date, the mechanism of GCRV entry into CIK Ctenopharyngon idellus kidney (CIK) cells remains poorly understood.

Results

Here, we present a study of the GCRV internalization mechanism in CIK cells. Our results indicated that GCRV infection was inhibited by chlorpromazine, the specific inhibitor for clathrin-mediated endocytosis. Colocalization of GCRV virions with endogenous clathrin was observed during early infection by confocal microscopy. Moreover, GCRV infection of CIK cells depended on acidification of the endosome. This was indicated by significant inhibition of viral infection following prophylactic treatment with the lysosomotropic drugs chloroquine or ammonium chloride. In addition, the disturbance of dynamin activity blocked GCRV entry, which confirmed the dynamin-dependent nature of clathrin-mediated endocytosis.

Conclusion

Our findings suggest that GCRV might enter CIK cells via clathrin-mediated endocytosis in a pH-dependent manner. Additionally, dynamin is critical for efficient viral entry.

Available only for authorised users

Wang Q, Cheng L, Liu J, Li Z, Xie S, De Silva SS. Freshwater aquaculture in PR China: trends and prospects. Rev Aquacult. 2015;7(4):283–302.

Xie B, Qin J, Yang H, Wang X, Wang Y-H, Li T-Y. Organic aquaculture in China: A review from a global perspective. Aquaculture. 2013;414:243–53.CrossRef

Jiang Y. Hemorrhagic disease of grass carp: status of outbreaks, diagnosis, surveillance, and research. Bamidgeh. 2009;61:188–197.

Qiu T, Lu R-H, Zhang J, Zhu Z-Y. Complete nucleotide sequence of the S10 genome segment of grass carp reovirus (GCRV). Dis Aquat Organ. 2001;44:69–74.CrossRefPubMed

Kim J, Tao Y, Reinisch KM, Harrison SC, Nibert ML. Orthoreovirus and Aquareovirus core proteins: conserved enzymatic surfaces, but not protein–protein interfaces. Virus Res. 2004;101:15–28.CrossRefPubMed

Mohd Jaafar F, Goodwin AE, Belhouchet M, Merry G, Fang Q, Cantaloube J-F, et al. Complete characterisation of the American grass carp reovirus genome (genus Aquareovirus: family Reoviridae) reveals an evolutionary link between aquareoviruses and coltiviruses. Virology. 2008;373:310–21.CrossRefPubMed

Attoui H, Fang Q, Jaafar FM, Cantaloube J-F, Biagini P, de Micco P, et al. Common evolutionary origin of aquareoviruses and orthoreoviruses revealed by genome characterization of Golden shiner reovirus, Grass carp reovirus, Striped bass reovirus and golden ide reovirus (genus Aquareovirus, family Reoviridae). J Gen Virol. 2002;83:1941–51.CrossRefPubMed

Zhang Q, Gui J-F. Virus genomes and virus-host interactions in aquaculture animals. Sci China Life Sci. 2015;58:156–69.CrossRefPubMed

Cheng L, Fang Q, Shah S, Atanasov IC, Zhou ZH. Subnanometer-resolution structures of the grass carp reovirus core and virion. J Mol Biol. 2008;382:213–22.PubMedCentralCrossRefPubMed

10.

Fan C, Shao L, Fang Q. Characterization of the nonstructural protein NS80 of grass carp reovirus. Arch Virol. 2010;155:1755–63.CrossRefPubMed

11.

Shao L, Guo H, Yan L-M, Liu H, Fang Q. Aquareovirus NS80 recruits viral proteins to its inclusions, and its C-terminal domain is the primary driving force for viral inclusion formation. PLoS One. 2013;8:e55334.PubMedCentralCrossRefPubMed

12.

Huang J, Li F, Wu J, Yang F. White spot syndrome virus enters crayfish hematopoietic tissue cells via clathrin-mediated endocytosis. Virology. 2015;486:35–43.CrossRefPubMed

13.

Bressanelli S, Stiasny K, Allison SL, Stura EA, Duquerroy S, Lescar J, et al. Structure of a flavivirus envelope glycoprotein in its low‐pH‐induced membrane fusion conformation. EMBO J. 2004;23:728–38.PubMedCentralCrossRefPubMed

14.

Han X, Bushweller JH, Cafiso DS, Tamm LK. Membrane structure and fusion-triggering conformational change of the fusion domain from influenza hemagglutinin. Nat Struct Mol Biol. 2001;8:715–20.CrossRef

15.

Skehel JJ, Wiley DC. Receptor binding and membrane fusion in virus entry: the influenza hemagglutinin. Annu Rev Biochem. 2000;69:531–69.CrossRefPubMed

16.

Corcoran JA, Duncan R. Reptilian reovirus utilizes a small type III protein with an external myristylated amino terminus to mediate cell-cell fusion. J Virol. 2004;78:4342–51.PubMedCentralCrossRefPubMed

17.

Smith AE, Helenius A. How viruses enter animal cells. Science. 2004;304:237–42.CrossRefPubMed

18.

Kumari S, Swetha M, Mayor S. Endocytosis unplugged: multiple ways to enter the cell. Cell Res. 2010;20:256–75.CrossRefPubMed

19.

Mayor S, Pagano RE. Pathways of clathrin-independent endocytosis. Nat Rev Mol Cell Biol. 2007;8:603–12.CrossRefPubMed

20.

van den Berg LM, Ribeiro CM, Zijlstra-Willems EM, de Witte L, Fluitsma D, Tigchelaar W, et al. Caveolin-1 mediated uptake via langerin restricts HIV-1 infection in human Langerhans cells. Retrovirology. 2014;11:3903.

21.

Zhang Y, Whittaker GR. Influenza entry pathways in polarized MDCK cells. Biochem Biophys Res Commun. 2014;450:234–9.PubMedCentralCrossRefPubMed

22.

Rossman JS, Leser GP, Lamb RA. Filamentous influenza virus enters cells via macropinocytosis. J Virol. 2012;86:10950–60.PubMedCentralCrossRefPubMed

23.

Quirin K, Eschli B, Scheu I, Poort L, Kartenbeck J, Helenius A. Lymphocytic choriomeningitis virus uses a novel endocytic pathway for infectious entry via late endosomes. Virology. 2008;378:21–33.CrossRefPubMed

24.

Schulz WL, Haj AK, Schiff LA. Reovirus uses multiple endocytic pathways for cell entry. J Virol. 2012;86:12665–75.PubMedCentralCrossRefPubMed

25.

Zuo W, Qian H, Xu Y, Du S, Yang X. A cell line derived from the kidney of grass carp. J Fish China. 1986;10:11–7.

26.

Wang T, Li J, Lu L. Quantitative in vivo and in vitro characterization of co-infection by two genetically distant grass carp reoviruses. J Gen Virol. 2013;94:1301–9.CrossRefPubMed

27.

Fang Q, Seng E, Ding Q, Zhang L. Characterization of infectious particles of grass carp reovirus by treatment with proteases. Arch Virol. 2008;153:675–82.CrossRefPubMed

28.

Macia E, Ehrlich M, Massol R, Boucrot E, Brunner C, Kirchhausen T. Dynasore, a cell-permeable inhibitor of dynamin. Dev Cell. 2006;10:839–50.CrossRefPubMed

29.

Nawa M, Takasaki T, Yamada K-I, Kurane I, Akatsuka T. Interference in Japanese encephalitis virus infection of Vero cells by a cationic amphiphilic drug, chlorpromazine. J Gen Virol. 2003;84:1737–41.CrossRefPubMed

30.

Wang L-H, Rothberg KG, Anderson R. Mis-assembly of clathrin lattices on endosomes reveals a regulatory switch for coated pit formation. J Cell Biol. 1993;123:1107–17.CrossRefPubMed

31.

Schnitzer JE, Oh P, Pinney E, Allard J. Filipin-sensitive caveolae-mediated transport in endothelium: reduced transcytosis, scavenger endocytosis, and capillary permeability of select macromolecules. J Cell Biol. 1994;127:1217–32.CrossRefPubMed

32.

He Y, Xu H, Yang Q, Xu D, Lu L. The use of an in vitro microneutralization assay to evaluate the potential of recombinant VP5 protein as an antigen for vaccinating against Grass carp reovirus. Virol J. 2011;8:132–8.PubMedCentralCrossRefPubMed

33.

Mercer J, Schelhaas M, Helenius A. Virus entry by endocytosis. Annu Rev Biochem. 2010;79:803–33.CrossRefPubMed

34.

Hernaez B, Alonso C. Dynamin-and clathrin-dependent endocytosis in African swine fever virus entry. J Virol. 2010;84:2100–9.PubMedCentralCrossRefPubMed

35.

Pickles RJ, McCarty D, Matsui H, Hart PJ, Randell SH, Boucher RC. Limited entry of adenovirus vectors into well-differentiated airway epithelium is responsible for inefficient gene transfer. J Virol. 1998;72:6014–23.PubMedCentralPubMed

36.

Pati S, Cavrois M, Guo H-G, Foulke JS, Kim J, Feldman RA, et al. Activation of NF-κB by the human herpesvirus 8 chemokine receptor ORF74: evidence for a paracrine model of Kaposi’s sarcoma pathogenesis. J Virol. 2001;75:8660–73.PubMedCentralCrossRefPubMed

37.

Kirchhausen T. Clathrin. Annu Rev Biochem. 2000;69:699–727.CrossRefPubMed

38.

Savarino A, Boelaert JR, Cassone A, Majori G, Cauda R. Effects of chloroquine on viral infections: an old drug against today’s diseases. Lancet Infect Dis. 2003;3:722–7.CrossRefPubMed

39.

Cassell S, Edwards J, Brown DT. Effects of lysosomotropic weak bases on infection of BHK-21 cells by Sindbis virus. J Virol. 1984;52:857–64.PubMedCentralPubMed

40.

Sever S, Damke H, Schmid SL. Dynamin: GTP controls the formation of constricted coated pits, the rate limiting step in clathrin-mediated endocytosis. J Cell Biol. 2000;150:1137–48.PubMedCentralCrossRefPubMed

41.

Sun X, Yau VK, Briggs BJ, Whittaker GR. Role of clathrin-mediated endocytosis during vesicular stomatitis virus entry into host cells. Virology. 2005;338:53–60.CrossRefPubMed

42.

Blanchard E, Belouzard S, Goueslain L, Wakita T, Dubuisson J, Wychowski C, et al. Hepatitis C virus entry depends on clathrin-mediated endocytosis. J Virol. 2006;80:6964–72.PubMedCentralCrossRefPubMed

43.

Pho M, Ashok A, Atwood WJ. JC virus enters human glial cells by clathrin-dependent receptor-mediated endocytosis. J Virol. 2000;74:2288–92.PubMedCentralCrossRefPubMed

44.

Marsh M, Helenius A. Virus entry into animal cells. Adv Virus Res. 1989;36:107–51.CrossRefPubMed

45.

Beer C, Andersen DS, Rojek A, Pedersen L. Caveola-dependent endocytic entry of amphotropic murine leukemia virus. J Virol. 2005;79:10776–87.PubMedCentralCrossRefPubMed

46.

Zou G, Fang Q. Study on replication and morphogenesis of the Grass Carp Reovirus (GCRV) in CIK cells. Virol Sin. 1999;15:188–92.

Title: Disruption of clathrin-dependent trafficking results in the failure of grass carp reovirus cellular entry
Authors: Hao Wang
Weisha Liu
Fei Yu
Liqun Lu
Publication date: 01-12-2016
Publisher: BioMed Central
Published in: Virology Journal / Issue 1/2016
Electronic ISSN: 1743-422X
DOI: https://doi.org/10.1186/s12985-016-0485-7

ACC 2024 Congress Coverage

Heart Failure Video

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Disruption of clathrin-dependent trafficking results in the failure of grass carp reovirus cellular entry

Abstract

Background

Results

Conclusion

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Background

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2016

Valproic acid ameliorates coxsackievirus-B3-induced viral myocarditis by modulating Th17/Treg imbalance

Identification of mumps virus protein and lipid composition by mass spectrometry

Occult Hepatitis B virus infection in previously screened, blood donors in Ile-Ife, Nigeria: implications for blood transfusion and stem cell transplantation

A heat-inactivated H7N3 vaccine induces cross-reactive cellular immunity in HLA-A2.1 transgenic mice

Circulation of avian paramyxoviruses in wild birds of Kazakhstan in 2002–2013

Molecular detection of Porcine astrovirus in Sichuan Province, China

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page