Characterization of an immunodominant epitope in the endodomain of the coronavirus membrane protein

Coronavirus; Endodomain; Immunodominant epitope; Membrane protein; M protein, Coronavirus; Article; Western blotting; Bagg albino mouse; Transmissible gastroenteritis virus; Animals; Antibodies, Monoclonal; Antibodies, Viral; Blotting, Western; Cell Line; Epitope Mapping; Fluorescent Antibody Technique; Immunodominant Epitopes; Mice, Inbred BALB C; Swine; Viral Matrix Proteins

Abstract :

[en] The coronavirus membrane (M) protein acts as a dominant immunogen and is a major player in virus assembly. In this study, we prepared two monoclonal antibodies (mAbs; 1C3 and 4C7) directed against the transmissible gastroenteritis virus (TGEV) M protein. The 1C3 and 4C7 mAbs both reacted with the native TGEV M protein in western blotting and immunofluorescence (IFA) assays. Two linear epitopes, 243YSTEART249 (1C3) and 243YSTEARTDNLSEQEKLLHMV262 (4C7), were identified in the endodomain of the TGEV M protein. The 1C3 mAb can be used for the detection of the TGEV M protein in different assays. An IFA method for the detection of TGEV M protein was optimized using mAb 1C3. Furthermore, the ability of the epitope identified in this study to stimulate antibody production was also evaluated. An immunodominant epitope in the TGEV membrane protein endodomain was identified. The results of this study have implications for further research on TGEV replication. © 2016 by the authors; licensee MDPI, Basel, Switzerland.

Disciplines :

Veterinary medicine & animal health

Author, co-author :

Dong, Hui ; Université de Liège - ULiège > Doct. sc. agro. & ingé. biol. (Paysage)

Zhang, X.; State Key Laboratory of Veterinary Biotechnology, Division of Swine Infectious Diseases, Harbin Veterinary Research Institute Chinese Academy of Agricultural Sciences, Harbin, China

Shi, H.; State Key Laboratory of Veterinary Biotechnology, Division of Swine Infectious Diseases, Harbin Veterinary Research Institute Chinese Academy of Agricultural Sciences, Harbin, China

Chen, J.; State Key Laboratory of Veterinary Biotechnology, Division of Swine Infectious Diseases, Harbin Veterinary Research Institute Chinese Academy of Agricultural Sciences, Harbin, China

Da, S.; State Key Laboratory of Veterinary Biotechnology, Division of Swine Infectious Diseases, Harbin Veterinary Research Institute Chinese Academy of Agricultural Sciences, Harbin, China

Zhu, Y.; State Key Laboratory of Veterinary Biotechnology, Division of Swine Infectious Diseases, Harbin Veterinary Research Institute Chinese Academy of Agricultural Sciences, Harbin, China

Li, F.; State Key Laboratory of Veterinary Biotechnology, Division of Swine Infectious Diseases, Harbin Veterinary Research Institute Chinese Academy of Agricultural Sciences, Harbin, China

Language :

English

Title :

Characterization of an immunodominant epitope in the endodomain of the coronavirus membrane protein

Publication date :

2016

Journal title :

Viruses

eISSN :

1999-4915

Publisher :

Multidisciplinary Digital Publishing Institute (MDPI), Switzerland

Volume :

Issue :

Peer reviewed :

Peer Reviewed verified by ORBi

Funders :

NSCF - National Natural Science Foundation of China

Funding text :

2015BAD12B02; 31572541, NSFC, National Natural Science Foundation of China; 31502092, NSFC, National Natural Science Foundation of China

Available on ORBi :

since 26 December 2020

Statistics

Number of views

58 (2 by ULiège)

Number of downloads

3 (2 by ULiège)

More statistics

Scopus citations^®

Scopus citations^®
without self-citations

OpenCitations

OpenAlex citations

Bibliography

De Groot, R.J.; Baker, S.G.; Baric, R.S.; Enjuanes, L.; Gorbalenya, A.E. Coronaviridae. In Virus Taxonomy: Ninth Report of the International Committee on Taxonomy of Viruses; King, A.M.Q., Adams, M.J., Carstens, E.B., Lefkowitz, E.J., Eds.; Elsevier Academic Press: San Diego, CA, USA, 2011; pp 774-796.
Reguera, J.; Santiago, C.; Mudgal, G.; Ordono, D.; Enjuanes, L.; Casasnovas, J.M. Structural bases of coronavirus attachment to host aminopeptidase N and its inhibition by neutralizing antibodies. PLoS Pathog. 2012, 8, e1002859.
Perlman, S.; Netland, J. Coronaviruses post-SARS: Update on replication and pathogenesis. Nat. Rev. Microbiol. 2009, 7, 439-450.
Lai, M.M.; Cavanagh, D. The molecular biology of coronaviruses. Adv. Virus Res. 1997, 48, 1-100.
Yang, D.; Leibowitz, J.L. The structure and functions of coronavirus genomic 3’ and 5 ends. Virus Res. 2015, 206, 120-133.
Jenwitheesuk, E.; Samudrala, R. Identifying inhibitors of the SARS coronavirus proteinase. Bioorg. Med. Chem. Lett. 2003, 13, 3989-3992.
Anand, K.; Ziebuhr, J.; Wadhwani, P.; Mesters, J.R.; Hilgenfeld, R. Coronavirus main proteinase (3CLpro) structure: Basis for design of anti-SARS drugs. Science 2003, 300, 1763-1767.
Yount, B.; Curtis, K.M.; Baric, R.S. Strategy for systematic assembly of large RNA and DNA genomes: Transmissible gastroenteritis virus model. J. Virol. 2000, 74, 10600-10611.
Sola, I.; Almazan, F.; Zuniga, S.; Enjuanes, L. Continuous and discontinuous RNA synthesis in coronaviruses. Annu. Rev. Virol. 2015, 2, 265-288.
Cruz, J.L.; Sola, I.; Becares, M.; Alberca, B.; Plana, J.; Enjuanes, L.; Zuñiga, S. Coronavirus gene 7 counteracts host defenses and modulates virus virulence. PLoS Pathog. 2011, 7, e1002090.
Zuniga, S.; Sola, I.; Moreno, J.L.; Sabella, P.; Plana-Duran, J.; Enjuanes, L. Coronavirus nucleocapsid protein is an RNA chaperone. Virology 2007, 357, 215-227.
Almazan, F.; Gonzalez, J.M.; Penzes, Z.; Izeta, A.; Calvo, E.; Plana-Duran, J.; Enjuanes, L. Engineering the largest RNA virus genome as an infectious bacterial artificial chromosome. Proc. Natl. Acad. Sci. USA 2000, 97, 5516-5521.
McBride, C.E.; Machamer, C.E. A single tyrosine in the severe acute respiratory syndrome coronavirus membrane protein cytoplasmic tail is important for efficient interaction with spike protein. J. Virol. 2010, 84, 1891-1901.
Swift, A.M.; Machamer, C.E. A Golgi retention signal in a membrane-spanning domain of coronavirus E1 protein. J. Cell Biol. 1991, 115, 19-30.
Armstrong, J.; Patel, S.; Riddle, P. Lysosomal sorting mutants of coronavirus E1 protein, a Golgi membrane protein. J. Cell Sci. 1990, 95, 191-197.
Locker, J.K.; Klumperman, J.; Oorschot, V.; Horzinek, M.C.; Geuze, H.J.; Rottier, P.J. The cytoplasmic tail of mouse hepatitis virus M protein is essential but not sufficient for its retention in the Golgi complex. J. Biol. Chem. 1994, 269, 28263-28269.
Hsieh, Y.C.; Li, H.C.; Chen, S.C.; Lo, S.Y. Interactions between M protein and other structural proteins of severe, acute respiratory syndrome-associated coronavirus. J. Biomed. Sci. 2008, 15, 707-717.
Siu, Y.L.; Teoh, K.T.; Lo, J.; Chan, C.M.; Kien, F.; Escriou, N.; Tsao, S.W.; Nicholls, J.M.; Altmeyer, R.; Peiris, J.S. The M, E, and N structural proteins of the severe acute respiratory syndrome coronavirus are required for efficient assembly, trafficking, and release of virus-like particles. J. Virol. 2008, 82, 11318-11330.
Tseng, Y.T.; Wang, S.M.; Huang, K.J.; Lee, A.I.; Chiang, C.C.; Wang, C.T. Self-assembly of severe acute respiratory syndrome coronavirus membrane protein. J. Biol. Chem. 2010, 285, 12862-12872.
Baudoux, P.; Carrat, C.; Besnardeau, L.; Charley, B.; Laude, H. Coronavirus pseudoparticles formed with mrecombinant M and E proteins induce alpha interferon synthesis by leukocytes. J. Virol. 1998, 72, 8636-8643.
Huang, Y.; Yang, Z.Y.; Kong, W.P.; Nabel, G.J. Generation of synthetic severe acute respiratory syndrome coronavirus pseudoparticles: Implications for assembly and vaccine production. J. Virol. 2004, 78, 12557-12565.
Mortola, E.; Roy, P. Efficient assembly and release of SARS coronavirus-like particles by a heterologous expression system. FEBS Lett. 2004, 576, 174-178.
De Haan, C.A.; Vennema, H.; Rottier, P.J. Assembly of the coronavirus envelope: Homotypic interactions between the M proteins. J. Virol. 2000, 74, 4967-4978.
Opstelten, D.J.; Raamsman, M.J.; Wolfs, K.; Horzinek, M.C.; Rottier, P.J. Envelope glycoprotein interactions in coronavirus assembly. J. Cell Biol. 1995, 131, 339-349.
De Haan, C.A.; Smeets, M.; Vernooij, F.; Vennema, H.; Rottier, P.J. Mapping of the coronavirus membrane protein domains involved in interaction with the spike protein. J. Virol. 1999, 73, 7441-7452.
Youn, S.; Collisson, E.W.; Machamer, C.E. Contribution of trafficking signals in the cytoplasmic tail of the infectious bronchitis virus spike protein to virus infection. J. Virol. 2005, 79, 13209-13217.
Nguyen, V.P.; Hogue, B.G. Protein interactions during coronavirus assembly. J. Virol. 1997, 71, 9278-9284.
Escors, D.; Ortego, J.; Laude, H.; Enjuanes, L. The membrane M protein carboxy terminus binds to transmissible gastroenteritis coronavirus core and contributes to core stability. J. Virol. 2001, 75, 1312-1324.
Rottier, P.; Brandenburg, D.; Armstrong, J.; van der Zeijst, B.; Warren, G. Assembly in vitro of a spanning membrane protein of the endoplasmic reticulum: The E1 glycoprotein of coronavirus mouse hepatitis virus A59. Proc. Natl. Acad. Sci. USA 1984, 81, 1421-1425.
Kapke, P.A.; Tung, F.Y.; Hogue, B.G.; Brian, D.A.; Woods, R.D.; Wesley, R. The amino-terminal signal peptide on the porcine transmissible gastroenteritis coronavirus matrix protein is not an absolute requirement for membrane translocation and glycosylation. Virology 1988, 165, 367-376.
Lopez, L.A.; Jones, A.; Arndt, W.D.; Hogue, B.G. Subcellular localization of SARS-CoV structural proteins. Adv. Exp. Med. Biol. 2006, 581, 297-300.
Nal, B.; Chan, C.; Kien, F.; Siu, L.; Tse, J.; Chu, K.; Kam, J.; Staropoli, I.; Crescenzo-Chaigne, B.; Escriou, N.; et al. Differential maturation and subcellular localization of severe acute respiratory syndrome coronavirus surface proteins S, M and E. J. Gen. Virol. 2005, 86, 1423-1434.
Escors, D.; Camafeita, E.; Ortego, J.; Laude, H.; Enjuanes, L. Organization of two transmissible gastroenteritis coronavirus membrane protein topologies within the virion and core. J. Virol. 2001, 75, 12228-12240.
Zhang, X.; Shi, H.; Chen, J.; Shi, D.; Li, C.; Feng, L. EF1A interacting with nucleocapsid protein of transmissible gastroenteritis coronavirus and plays a role in virus replication. Vet. Microbiol. 2014, 172, 443-448.
Kohler, G.; Milstein, C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975, 256, 495-497.
Jungmann, A.; Nieper, H.; Muller, H. Apoptosis is induced by infectious bursal disease virus replication in productively infected cells as well as in antigen-negative cells in their vicinity. J. Gen. Virol. 2001, 82, 1107-1115.
Wang, X.; Qiu, H.; Zhang, M.; Cai, X.; Qu, Y.; Hu, D.; Zhao, X.; Zhou, E.; Liu, S.; Xiao, Y. Distribution of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) in different stages of gestation sows: HP-PRRSV distribution in gestation sows. Vet. Immunol. Immunopathol. 2015, 166, 88-94.
Biasini, M.; Bienert, S.; Waterhouse, A.; Arnold, K.; Studer, G.; Schmidt, T.; Kiefer, F.; Gallo Cassarino, T.; Bertoni, M.; Bordoli, L. SWISS-MODEL: Modelling protein tertiary and quaternary structure using evolutionary information. Nucleic Acids Res. 2014, 42, 252-258.
Love, M.L.; Szebenyi, D.M.; Kriksunov, I.A.; Thiel, D.J.; Munshi, C.; Graeff, R.; Lee, H.C.; Hao, Q. ADP-ribosyl cyclase; crystal structures reveal a covalent intermediate. Structure 2004, 12, 477-486.
Yano, A.; Miwa, Y.; Kanazawa, Y.; Ito, K.; Makino, M.; Imai, S.; Hanada, N.; Nisizawa, T. A novel method for enhancement of peptide vaccination utilizing T-cell epitopes from conventional vaccines. Vaccine 2013, 31, 1510-1515.
Kuo, L.; Hurst-Hess, K.R.; Koetzner, C.A.; Masters, P.S. Analyses of coronavirus assembly interactions with interspecies membrane and nucleocapsid protein chimeras. J. Virol. 2016, 90, 4357-4368.
Zhang, Z.; Chen, J.; Shi, H.; Chen, X.; Shi, D.; Feng, L.; Yang, B. Identification of a conserved linear B-cell epitope in the M protein of porcine epidemic diarrhea virus. Virol. J. 2012, 9.
Xing, J.; Liu, S.; Han, Z.; Shao, Y.; Li, H.; Kong, X. Identification of a novel linear B-cell epitope in the M protein of avian infectious bronchitis coronaviruses. J. Microbiol. 2009, 47, 589-599.
He, Y.; Zhou, Y.; Siddiqui, P.; Niu, J.; Jiang, S. Identification of immunodominant epitopes on the membrane protein of the severe acute respiratory syndrome-associated coronavirus. J. Clin. Microbiol. 2005, 43, 3718-3726.
Laviada, M.D.; Videgain, S.P.; Moreno, L.; Alonso, F.; Enjuanes, L.; Escribano, J.M. Expression of swine transmissible gastroenteritis virus envelope antigens on the surface of infected cells: Epitopes externally exposed. Virus Res. 1990, 16, 247-254.
De Diego, M.; Laviada, M.D.; Enjuanes, L.; Escribano, J.M. Epitope specificity of protective lactogenic immunity against swine transmissible gastroenteritis virus. J. Virol. 1992, 66, 6502-6508.
Rodak, L.; Smid, B.; Nevorankova, Z.; Valicek, L.; Smitalova, R. Use of monoclonal antibodies in blocking ELISA detection of transmissible gastroenteritis virus in faeces of piglets. J. Vet. Med. B Infect. Dis. Vet. Public Health 2005, 52, 105-111.
Rottier, P.J.; Welling, G.W.; Welling-Wester, S.; Niesters, H.G.; Lenstra, J.A.; van der Zeijst, B.A. Predicted membrane topology of the coronavirus protein E1. Biochemistry 1986, 25, 1335-1339.
Neuman, B.W.; Kiss, G.; Kunding, A.H.; Bhella, D.; Baksh, M.F.; Connelly, S.; Droese, B.; Klaus, J.P.; Makino, S.; Sawicki, S.G. A structural analysis of M protein in coronavirus assembly and morphology. J. Struct. Biol. 2011, 174, 11-22.
Kuo, L.; Masters, P.S. Evolved variants of the membrane protein can partially replace the envelope protein in murine coronavirus assembly. J. Virol. 2010, 84, 12872-12885.
Kuo, L.; Masters, P.S. Genetic evidence for a structural interaction between the carboxy termini of the membrane and nucleocapsid proteins of mouse hepatitis virus. J. Virol. 2002, 76, 4987-4999.
Hurst, K.R.; Kuo, L.; Koetzner, C.A.; Ye, R.; Hsue, B.; Masters, P.S. A major determinant for membrane protein interaction localizes to the carboxy-terminal domain of the mouse coronavirus nucleocapsid protein. J. Virol. 2005, 79, 13285-13297.
Verma, S.; Lopez, L.A.; Bednar, V.; Hogue, B.G. Importance of the penultimate positive charge in mouse hepatitis coronavirus A59 membrane protein. J. Virol. 2007, 81, 5339-5348.
Luo, H.; Wu, D.; Shen, C.; Chen, K.; Shen, X.; Jiang, H. Severe acute respiratory syndrome coronavirus membrane protein interacts with nucleocapsid protein mostly through their carboxyl termini by electrostatic attraction. Int. J. Biochem. Cell Biol. 2006, 38, 589-599.
He, R.; Leeson, A.; Ballantine, M.; Andonov, A.; Baker, L.; Dobie, F.; Li, Y.; Bastien, N.; Feldmann, H.; Strocher, U. Characterization of protein-protein interactions between the nucleocapsid protein and membrane protein of the SARS coronavirus. Virus Res. 2004, 105, 121-125.
Fang, X.; Ye, L.; Timani, K.A.; Li, S.; Zen, Y.; Zhao, M.; Zheng, H.; Wu, Z. Peptide domain involved in the interaction between membrane protein and nucleocapsid protein of SARS-associated coronavirus. J. Biochem. Mol. Biol. 2005, 38, 381-385.
Anton, I.M.; Gonzalez, S.; Bullido, M.J.; Corsin, M.; Risco, C.; Langeveld, J.P.; Enjuanes, L. Cooperation between transmissible gastroenteritis coronavirus (TGEV) structural proteins in the in vitro induction of virus-specific antibodies. Virus Res. 1996, 46, 111-124.
Kirchdoerfer, R.N.; Cottrell, C.A.; Wang, N.; Pallesen, J.; Yassine, H.M.; Turner, H.L.; Corbett, K.S.; Graham, B.S.; McLellan, J.S.; Ward, A.B. Pre-fusion structure of a human coronavirus spike protein. Nature 2016, 531, 118-121.
Belouzard, S.; Millet, J.K.; Licitra, B.N.; Whittaker, G.R. Mechanisms of coronavirus cell entry mediated by the viral spike protein. Viruses 2012, 4, 1011-1033.
Gao, J.; Lu, G.; Qi, J.; Li, Y.; Wu, Y.; Deng, Y.; Geng, H.; Li, H.; Wang, Q.; Xiao, H. Structure of the fusion core and inhibition of fusion by a heptad repeat peptide derived from the S protein of Middle East respiratory syndrome coronavirus. J. Virol. 2013, 87, 13134-13140.
Wicht, O.; Burkard, C.; de Haan, C.A.; van Kuppeveld, F.J.; Rottier, P.J.; Bosch, B.J. Identification and characterization of a proteolytically primed form of the murine coronavirus spike proteins after fusion with the target cell. J. Virol. 2014, 88, 4943-4952.
Hofmann, H.; Hattermann, K.; Marzi, A.; Gramberg, T.; Geier, M.; Krumbiegel, M.; Kuate, S.; Uberla, K.; Niedrig, M.; Pohlmann, S. S protein of severe acute respiratory syndrome-associated coronavirus mediates entry into hepatoma cell lines and is targeted by neutralizing antibodies in infected patients. J. Virol. 2004, 78, 6134-6142.
Chen, Z.; Zhang, L.; Qin, C.; Ba, L.; Yi, C.E.; Zhang, F.; Wei, Q.; He, T.; Yu, W.; Yu, J. Recombinant modified vaccinia virus Ankara expressing the spike glycoprotein of severe acute respiratory syndrome coronavirus induces protective neutralizing antibodies primarily targeting the receptor binding region. J. Virol. 2005, 79, 2678-2688.
Risco, C.; Anton, I.M.; Sune, C.; Pedregosa, A.M.; Martín-Alonso, J.M.; Parra, F.; Carrascosa, J.L.; Enjuanes, L. Membrane protein molecules of transmissible gastroenteritis coronavirus also expose the carboxy-terminal region on the external surface of the virion. J. Virol. 1995, 69, 5269-5277.
Fan, J.H.; Zuo, Y.Z.; Shen, X.Q.; Gu, W.Y.; Di, J.M. Development of an enzyme-linked immunosorbent assay for the monitoring and surveillance of antibodies to porcine epidemic diarrhea virus based on a recombinant membrane protein. J. Virol. Methods 2015, 225, 90-94.