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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Vp28 of Shrimp White Spot Syndrome Virus Is Involved in the Attachment and Penetration into Shrimp Cells

  • Yi, Guohua (Department of Virology, College of Life Sciences, Wuhan University) ;
  • Wang, Zhimin (Department of Virology, College of Life Sciences, Wuhan University) ;
  • Qi, Yipeng (Department of Virology, College of Life Sciences, Wuhan University) ;
  • Yao, Lunguang (Department of Virology, College of Life Sciences, Wuhan University) ;
  • Qian, Juan (Department of Virology, College of Life Sciences, Wuhan University) ;
  • Hu, Longbo (Department of Virology, College of Life Sciences, Wuhan University)
  • Published : 2004.11.30
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Abstract

White spot disease (WSD) is caused by the white spot syndrome virus (WSSV), which results in devastating losses to the shrimp farming industry around the world. However, the mechanism of virus entry and spread into the shrimp cells is unknown. A binding assay in vitro demonstrated VP28-EGFP (envelope protein VP28 fused with enhanced green fluorescence protein) binding to shrimp cells. This provides direct evidence that VP28-EGFP can bind to shrimp cells at pH 6.0 within 0.5 h. However, the protein was observed to enter the cytoplasm 3 h post-adsorption. Meanwhile, the plaque inhibition test showed that the polyclonal antibody against VP28 (a major envelope protein of WSSV) could neutralize the WSSV and block an infection with the virus. The result of competition ELISA further confirmed that the envelope protein VP28 could compete with WSSV to bind to shrimp cells. Overall, VP28 of the WSSV can bind to shrimp cells as an attachment protein, and can help the virus enter the cytoplasm.

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References

  1. Blissard, G. W. and Wenz, J. R. (1992) Baculovirus gp64 envelope glycoprotein is sufficient to mediate pH dependent membrane fusion. J. Virol. 66, 6829-6835.
  2. Chen, L. L., Leu, J. H., Huang, C. J., Chou, C. M., Chen, S. M., Wang, C. H., Lo, C. F. and Kou, G. H. (2002) Identification of a nucleocapsid protein (VP35) gene of shrimp white spot syndrome virus and characterization of the motif important for targeting VP35 to the nuclei of transfected insect cells. Virology 293, 44-53. https://doi.org/10.1006/viro.2001.1273
  3. Chen Y. P., Maguire T. and Marks R. M. (1996) Demonstration of binding of dengue virus envelope protein to target cells. J. Virol. 70, 8765-8772.
  4. Chen, X. F., Chen, C., Wu, D. H., Huai, H. and Chi, X. C. (1997) A new baculovirus of cultured shrimp. Sci. China Ser. C 40, 630-635. https://doi.org/10.1007/BF02882693
  5. Fayolle, J., Verrier, B., Buckland, R. and Wild, T. F. (1999) Charecterization of a natural mutation in an antigenic site on the fusion protein of measles virus that is involved in neutralization. J. Virol. 73, 787-790.
  6. Flegel, T. W. (1997) Special topic review: Major viral diseases of the black tiger prawn (Penaeus monodon) in Thailand. Wld. J. Microbiol. Biotechnol. 13, 433-442. https://doi.org/10.1023/A:1018580301578
  7. Francki, R. I. B., Fauquet, C. M., Knudson, D. L. and Brown, F. (1991) Classification and nomenclature of viruses. Fifth report of the International Commitee on Taxonomy of viruses. Arch. Virol. 2, 1450.
  8. Galmiche, M. C., Goenaga, J., Wittek, R. and Rindisbacher, L. (1999) Neutralizing and protective antibodies directed against vaccinia virus envelope antigens. Virology 254, 71-80. https://doi.org/10.1006/viro.1998.9516
  9. Gong, S. C., Lai, C. F. and Esteban, M. (1990) Viccinia virus induces cell fusion at acid pH and this activity is mediated by the N-terminus of the 14-kDa virus envelope protein. Virology 178, 81-91. https://doi.org/10.1016/0042-6822(90)90381-Z
  10. Guo, J. T. and Pugh, J. C. (1997) Topology of the large envelope protein of duck hepatitis B virus suggests a mechanism for membrane translocation during particle morphogenisis. J. Virol. 71, 1107-1114.
  11. Lai, C. F., Gong, S. C. and Esteban,. (1991) The 32-kilodalton envelope protein of vaccinia virus synthesized in Escherichia coli binds with specificity to cell surfaces. J. Virol. 65, 499- 504.
  12. Lambert, D. M., Barney, S., Lambert, A. L., Guthrie, K., Medinas, R., Davis, D. E., Bucy, T., Erickson, J., Merutka, G., and Petteway, S. R., Jr (1996) Peptides from conserved regions of powamyxovirus fusion (F) proteins are potent inhibitors of viral fusion. Proc. Natl. Acad. Sci. USA 93, 2186-2191. https://doi.org/10.1073/pnas.93.5.2186
  13. Li, L. and Qi, Y. (2002) A novel amino acid position in hemagglutinin glycoprotein of measles virus is responsible for hemadsorption and CD46 binding. Arch. Virol. 147, 775-786. https://doi.org/10.1007/s007050200025
  14. Liu, W. J., Yu, H. T., Peng, S. E., Chang, Y. S., Pien, H. W., Lin, C. J., Huang, C. J., Tsai, M. F., Huang, C. J., Wang, C. H., Lin, J. Y., Lo, C. F. and Kou, G. H. (2001) Cloning, characterization, and phylogenetic analysis of a shrimp white spot syndrome virus gene that encodes a protein kinase. Virology 289, 362-377. https://doi.org/10.1006/viro.2001.1091
  15. Lo, C. F., Ho, C. H., Peng, S. E., Chen, C. H., Hsu, H. C., Chiu, Y. L., Chang, C. F., Liu, K. F., Su, M. S., Wang, C. H. and Kou, G. H. (1996) White spot syndrome baculovirus (WSBV) detected in cultured and captured shrimp, crabs and other arthropods. Dis. Aquat. Org. 27, 215-225. https://doi.org/10.3354/dao027215
  16. Mass, J. S., Rodriguez, J. F. and Esteban, M. (1990) Structural and functional characterization of a cell surface binding protein of vaccinia virus. J. Biol. Chem. 265, 1569-1577.
  17. Molesworth, S. J., Lake, C. M., Borza, C. M., Turk, S. M. and Hutt-Fletcher, L. M. (2000) Epstein-Barr virus gH is essential for penetration of B cells but also plays a role in attachment of virus to epithelial Cells. J. Virol. 74, 6324-6332. https://doi.org/10.1128/JVI.74.14.6324-6332.2000
  18. Murphy, F. A., Fauquet, C. M., Mayo, M. A., Jarvis, A. W., Ghabrial, S. A., Summers, M. D., Martelli, G. P. and Bishop, D. H. L. (1995) Classification and nomenclature of viruses; in sixth report of the international committee on taxonomy of viruses. Arch. Virol. 10, 428-433.
  19. Ottmar, H., Dieter, M., Michael, B., Thomas, P., Axel, R., Paul, D. B., Myra, O. M., Roland, W. and Josef, S. (1999) Specific binding of recombinant foamy virus envelope protein to host cells correlates with susceptibility to infection. Virology 255, 228-236. https://doi.org/10.1006/viro.1998.9570
  20. Peeters, B., de Wind, N., Hooisma, M., Wagenaar, F., Gielkens, A. and Moormann, R. (1992) Pseudorabies virus envelope glycoproteins gp50 and gII are essential for virus penetration, but only gII is involved in membrane fusion. J. Virol. 66, 894- 905.
  21. Ramsey, I. K., Spibey, N. and Jarrett, O. (1998) The receptor binding site of feline leukemia virus surface glycoprotein is distinct from the site involed in virus neutralization. J. Virol. 72, 3268-3277.
  22. Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, USA.
  23. Sunyach, C., Rollier, C., Robaczewska, M., Borel, C., Barraud, L., Kay, A., Trepo, C., will, H. and Cova, L. (1999) Residues critical for duck hepatitis B virus neutralization are involved in host cell interaction. J. Virol. 73, 2569-2575.
  24. Tyler, K. L. and Fields, B. N. (1996) Pathogenesis of viral infections; in Fields Virology, Fields, B. N., Knipe, D. M., Howley, P. M. et al. (eds), pp. 173-217, Lippincott Raven, Philadelphia, USA.
  25. Van Hulten, M. C., Reijns, M., Vermeesch, A. M., Zandbergen, F. and Vlak, J. M. (2002) Identification of VP19 and VP15 of white spot syndrome virus (WSSV) and glycosylation status of the WSSV major structural proteins. J. Gen. Virol. 83, 257-265.
  26. Van Hulten, M. C., Goldbach, R. W. and Vlak, J. M., (2000b) Three functionally diverged major structural proteins of white spot syndrome virus evolved by gene duplication. J. Gen. Virol. 81, 2525-2529.
  27. Van Hulten, M. C., Westenberg, M., Goodall, S. D. and Vlak, J. M. (2000a) Identification of two major virion protein genes of white spot syndrome virus of shrimp. Virology 266, 227-236. https://doi.org/10.1006/viro.1999.0088
  28. Van Hulten, M. C., Witteveldt, J., Peters, S., Kloosterboer, N., Tarchini, R., Fiers, M., Sandbrink, H., Lankhorst, R. K. and Vlak, J. M. (2001a) The white spot syndrome virus DNA genome sequence. Virology 286, 7-22. https://doi.org/10.1006/viro.2001.1002
  29. Van Hulten, M. C., Witteveldt, J., Snippe, M. and Vlak, J. M. (2001b) White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp. Virology 285, 228-233. https://doi.org/10.1006/viro.2001.0928
  30. Volkman, L. E. and Goldsmith, P. A. (1985) Mechanism of neutralization of budded Autographa California nuclear polyhedrosis virus by a monoclonal antibody: Inhibition of entry by adsorptive endocytosis. Virology 143, 185-195. https://doi.org/10.1016/0042-6822(85)90107-2
  31. Witteveldt, J, Van Hulten, M. C. and Vlak, J. M. (2001) Identification and phylogeny of a non-specific endonuclease gene of white spot syndrome virus of shrimp. Virus Genes 23, 331-337. https://doi.org/10.1023/A:1012529524663
  32. Yang, F., He, J., Lin, X. H., Li, Q., Pan, D., Zhang, X. B. and Xu, X. (2001) Complete genome sequence of the shrimp white spot bacilliform virus. J. Virol. 75, 11811-11820. https://doi.org/10.1128/JVI.75.23.11811-11820.2001
  33. Yi, G., Qian, J., Wang, Z. and Qi, Y. (2003) A phage-displayed peptide can inhibit infection by white spot syndrome virus of shrimp. J. Gen. Virol. 84, 2545-2553. https://doi.org/10.1099/vir.0.19001-0
  34. Zhang, X., Huang, C., Xu, X. and Hew, C. L. (2002) Transcription and identification of an envelope protein gene (p22) from shrimp white spot syndrome virus. J. Gen. Virol. 83, 471-477.

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  7. Analysis of white spot syndrome virus envelope protein complexome by two-dimensional blue native/SDS PAGE combined with mass spectrometry vol.156, pp.7, 2011, https://doi.org/10.1007/s00705-011-0954-7
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  11. Binding of shrimp cellular proteins to Taura syndrome viral capsid proteins VP1, VP2 and VP3 vol.122, pp.1-2, 2006, https://doi.org/10.1016/j.virusres.2006.06.013
  12. Reprint of: Virus-binding proteins and their roles in shrimp innate immunity vol.34, pp.4, 2013, https://doi.org/10.1016/j.fsi.2013.02.013
  13. An Elegant Analysis of White Spot Syndrome Virus Using a Graphene Oxide/Methylene Blue based Electrochemical Immunosensor Platform vol.7, 2017, https://doi.org/10.1038/srep46169
  14. Localization of VP28 on the baculovirus envelope and its immunogenicity against white spot syndrome virus in Penaeus monodon vol.391, pp.2, 2009, https://doi.org/10.1016/j.virol.2009.06.017
  15. Involvement of WSSV–shrimp homologs in WSSV infectivity in kuruma shrimp: Marsupenaeus japonicus vol.88, pp.2, 2010, https://doi.org/10.1016/j.antiviral.2010.08.017
  16. White Spot Syndrome Virus in cultured shrimp: A review vol.38, pp.13, 2007, https://doi.org/10.1111/j.1365-2109.2007.01827.x
  17. Development of paratransgenicArtemiaas a platform for control of infectious diseases in shrimp mariculture vol.108, pp.3, 2010, https://doi.org/10.1111/j.1365-2672.2009.04479.x
  18. VP37 of white spot syndrome virus interact with shrimp cells vol.48, pp.1, 2009, https://doi.org/10.1111/j.1472-765X.2008.02482.x
  19. Diversity and multiple functions of lectins in shrimp immunity vol.39, pp.1-2, 2013, https://doi.org/10.1016/j.dci.2012.04.009
  20. Counter-Insurgents of the Blue Revolution? Parasites and Diseases Affecting Aquaculture and Science vol.100, pp.6, 2014, https://doi.org/10.1645/14-605.1
  21. Passive immunity in tiger shrimp (Penaeus monodon) fed with monoclonal antibody to white spot syndrome virus vol.22, pp.2, 2014, https://doi.org/10.1007/s10499-013-9714-x
  22. A C-type lectin is involved in the innate immune response of Chinese white shrimp vol.27, pp.4, 2009, https://doi.org/10.1016/j.fsi.2009.07.011
  23. A vector that expresses VP28 of WSSV can protect red swamp crayfish from white spot disease vol.36, pp.2, 2012, https://doi.org/10.1016/j.dci.2011.08.009
  24. Molecular Mechanisms of White Spot Syndrome Virus Infection and Perspectives on Treatments vol.8, pp.1, 2016, https://doi.org/10.3390/v8010023
  25. Oral Administration of Bacterially Expressed VP28dsRNA to Protect Penaeus monodon from White Spot Syndrome Virus vol.10, pp.3, 2008, https://doi.org/10.1007/s10126-007-9057-6
  26. Epitope analysis of white spot syndrome virus of Penaeus monodon by in vivo neutralization assay employing a panel of monoclonal antibodies vol.30, pp.4-5, 2011, https://doi.org/10.1016/j.fsi.2011.01.023
  27. Sequence Analysis of cDNAs Encoding the Heavy and Light Chain Variable Regions of a WSSV-Neutralizing Monoclonal Antibody vol.27, pp.6, 2008, https://doi.org/10.1089/hyb.2008.0055
  28. Blocking the large extracellular loop (LEL) domain of FcTetraspanin-3 could inhibit the infection of white spot syndrome virus (WSSV) in Chinese shrimp, Fenneropenaeus chinensis vol.32, pp.6, 2012, https://doi.org/10.1016/j.fsi.2012.02.022
  29. Neutralizing assay of monoclonal antibodies against white spot syndrome virus (WSSV) vol.272, pp.1-4, 2007, https://doi.org/10.1016/j.aquaculture.2007.07.229
  30. Protection of Procambarus clarkii against white spot syndrome virus using recombinant subunit injection vaccine expressed in Pichia pastoris vol.72, pp.5, 2006, https://doi.org/10.1111/j.1444-2906.2006.01250.x
  31. Antiviral immunity in crustaceans vol.27, pp.2, 2009, https://doi.org/10.1016/j.fsi.2009.02.009
  32. A review on the morphology, molecular characterization, morphogenesis and pathogenesis of white spot syndrome virus vol.31, pp.1, 2008, https://doi.org/10.1111/j.1365-2761.2007.00877.x
  33. Oral delivery of DNA vaccine encoding VP28 against white spot syndrome virus in crayfish by attenuated Salmonella typhimurium vol.27, pp.7, 2009, https://doi.org/10.1016/j.vaccine.2008.11.075
  34. Preparation of transgenic Dunaliella salina for immunization against white spot syndrome virus in crayfish vol.159, pp.3, 2014, https://doi.org/10.1007/s00705-013-1856-7
  35. Injected phage-displayed-VP28 vaccine reduces shrimp Litopenaeus vannamei mortality by white spot syndrome virus infection vol.55, 2016, https://doi.org/10.1016/j.fsi.2016.05.027
  36. Inactivation of White Spot Syndrome Virus (WSSV) by normal rabbit serum: Implications for the role of the envelope protein VP28 in WSSV infection of shrimp vol.118, pp.1-2, 2006, https://doi.org/10.1016/j.virusres.2005.11.011
  37. Identification of the interaction domains of white spot syndrome virus envelope proteins VP28 and VP24 vol.200, 2015, https://doi.org/10.1016/j.virusres.2015.01.017
  38. Reprint of “Evolution of specific immunity in shrimp – A vaccination perspective against white spot syndrome virus” vol.48, pp.2, 2015, https://doi.org/10.1016/j.dci.2014.07.016
  39. Oral vaccination with envelope protein VP28 against white spot syndrome virus in Procambarus clarkii using Bacillus subtilis as delivery vehicles vol.46, pp.5, 2008, https://doi.org/10.1111/j.1472-765X.2008.02355.x
  40. Bacillus subtilisspores expressing the VP28 antigen: a potential oral treatment to protectLitopenaeus vannameiagainst white spot syndrome vol.358, pp.2, 2014, https://doi.org/10.1111/1574-6968.12546
  41. Characterization and diagnostic use of a monoclonal antibody for VP28 envelope protein of white spot syndrome virus vol.26, pp.4, 2011, https://doi.org/10.1007/s12250-011-3202-0
  42. White spot syndrome virus (WSSV) inactivation in Penaeus japonicus using purified monoclonal antibody targeting viral envelope protein vol.269, pp.1-4, 2007, https://doi.org/10.1016/j.aquaculture.2007.03.036
  43. Virus-binding proteins and their roles in shrimp innate immunity vol.33, pp.6, 2012, https://doi.org/10.1016/j.fsi.2012.09.017
  44. Lateral flow assay for rapid detection of white spot syndrome virus (WSSV) using a phage-displayed peptide as bio-recognition probe vol.101, pp.11, 2017, https://doi.org/10.1007/s00253-017-8232-6
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  47. Cloning and characterization of three novel WSSV recognizing lectins from shrimp Marsupenaeus japonicus vol.28, pp.4, 2010, https://doi.org/10.1016/j.fsi.2009.12.015
  48. Effect of high water temperature on mortality, immune response and viral replication of WSSV-infected Marsupenaeus japonicus juveniles and adults vol.305, pp.1-4, 2010, https://doi.org/10.1016/j.aquaculture.2010.04.024
  49. In vivo assessment for oral delivery of Bacillus subtilis harboring a viral protein (VP28) against white spot syndrome virus in Litopenaeus vannamei vol.322-323, 2011, https://doi.org/10.1016/j.aquaculture.2011.09.036
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  52. Double-stranded RNA-mediated silencing of the white spot syndrome virus VP28 gene in kuruma shrimp, Marsupenaeus japonicus vol.42, pp.8, 2011, https://doi.org/10.1111/j.1365-2109.2010.02703.x
  53. Proteomic analyses of the shrimp white spot syndrome virus vol.23, pp.3, 2008, https://doi.org/10.1007/s12250-008-2924-0
  54. White spot syndrome virus: an overview on an emergent concern vol.41, pp.6, 2010, https://doi.org/10.1051/vetres/2010015
  55. Comparative efficacy of double-stranded RNAs targeting WSSV structural and nonstructural genes in controlling viral multiplication in Penaeus monodon vol.157, pp.5, 2012, https://doi.org/10.1007/s00705-012-1258-2
  56. Silencing shrimp white spot syndrome virus (WSSV) genes by siRNA vol.73, pp.2, 2007, https://doi.org/10.1016/j.antiviral.2006.08.007
  57. TAT-mediated oral subunit vaccine against white spot syndrome virus in crayfish vol.181, pp.1, 2012, https://doi.org/10.1016/j.jviromet.2012.01.011
  58. Protection of Penaeus monodon from Infection of White spot syndrome virus by DNA Construct Expressing Long Hairpin-RNA Against ICP11 Gene vol.21, pp.2, 2010, https://doi.org/10.1007/s13337-011-0024-5
  59. DNA constructs expressing long-hairpin RNA (lhRNA) protect Penaeus monodon against White Spot Syndrome Virus vol.27, pp.29, 2009, https://doi.org/10.1016/j.vaccine.2009.04.011
  60. Evolution of specific immunity in shrimp – A vaccination perspective against white spot syndrome virus vol.46, pp.2, 2014, https://doi.org/10.1016/j.dci.2014.04.013
  61. Protection of Procambarus clarkii against white spot syndrome virus using inactivated WSSV vol.26, pp.5, 2009, https://doi.org/10.1016/j.fsi.2009.02.022
  62. Three members of Ras GTPase superfamily are response to white spot syndrome virus challenge in Marsupenaeus japonicus vol.55, 2016, https://doi.org/10.1016/j.fsi.2016.06.038
  63. Interaction between white spot syndrome virus VP26 and hemocyte membrane of shrimp, Fenneropenaeus chinensis vol.314, pp.1-4, 2011, https://doi.org/10.1016/j.aquaculture.2011.01.023
  64. Virulence status, viral accommodation and structural protein profiles of white spot syndrome virus isolates in farmedPenaeus monodonfrom the southeast coast of India vol.40, pp.2, 2009, https://doi.org/10.1111/j.1365-2109.2008.02071.x
  65. Development and validation of a quantitative real-time polymerase chain assay for universal detection of the White Spot Syndrome Virus in marine crustaceans vol.10, pp.1, 2013, https://doi.org/10.1186/1743-422X-10-186
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