1887

Abstract

Inhibition of the host melanization reaction, activated by the prophenoloxidase activating (proPO) system, is one of the crucial evasion strategies of pathogens. Recently, the shrimp pathogen, white spot syndrome virus (WSSV), was found to inhibit melanization in the shrimp Penaeus monodon. The viral protein WSSV453 was previously shown to interact with PO-activating enzyme 2 (PmPPAE2) and reported to be involved in suppressing the shrimp melanization response after WSSV infection. Here, we characterized how WSSV453 inhibits melanization. WSSV453 is a non-structural viral protein, which was first detected in shrimp haemocytes at 6 hours post-infection (hpi) by WSSV and in shrimp plasma at 24 hpi. We produced recombinant proteins for three components of the P. monodon proPO system: PmproPPAE2, PmproPO1 and PmproPO2. Functional assays showed that active PmPPAE2 processed PmproPO1 and 2 to produce functional PO. Incubation of WSSV453 with PmproPPAE2 dose-dependently reduced PmPPAE2 activity toward PmPO1 or PmPO2. In contrast, WSSV453 had no effect on activated PmPPAE2. The addition of active PmPPAE2 to WSSV-infected shrimp plasma at day 2 post-infection also rescued PO activity. Taken together, these results indicate that the anti-melanization activity of WSSV is due to WSSV453, which interacts with PmproPPAE2 and interferes with its activation to active PmPPAE2.

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2017-04-28
2019-10-20
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References

  1. Finlay BB, McFadden G. Anti-immunology: evasion of the host immune system by bacterial and viral pathogens. Cell 2006; 124: 767– 782 [Crossref]
    [Google Scholar]
  2. Cerenius L, Lee BL, Söderhäll K. The proPO-system: pros and cons for its role in invertebrate immunity. Trends Immunol 2008; 29: 263– 271 [Crossref]
    [Google Scholar]
  3. Kanost MR, Gorman MJ. Phenoloxidases in insect immunity. Insect Immunology 2008; 69– 96 [Crossref]
    [Google Scholar]
  4. Amparyup P, Charoensapsri W, Tassanakajon A. Prophenoloxidase system and its role in shrimp immune responses against major pathogens. Fish Shellfish Immunol 2013; 34: 990– 1001 [Crossref]
    [Google Scholar]
  5. Söderhäll K, Cerenius L. Role of the prophenoloxidase-activating system in invertebrate immunity. Curr Opin Immunol 1998; 10: 23– 28 [CrossRef] [PubMed]
    [Google Scholar]
  6. Cerenius L, Kawabata S, Lee BL, Nonaka M, Söderhäll K. Proteolytic cascades and their involvement in invertebrate immunity. Trends Biochem Sci 2010; 35: 575– 583 [Crossref]
    [Google Scholar]
  7. Amparyup P, Charoensapsri W, Tassanakajon A. Two prophenoloxidases are important for the survival of Vibrio harveyi challenged shrimp Penaeus monodon. Dev Comp Immunol 2009; 33: 247– 256 [CrossRef] [PubMed]
    [Google Scholar]
  8. Binggeli O, Neyen C, Poidevin M, Lemaitre B. Prophenoloxidase activation is required for survival to microbial infections in Drosophila. PLoS Pathog 2014; 10: e1004067 [Crossref]
    [Google Scholar]
  9. Nappi AJ, Christensen BM. Melanogenesis and associated cytotoxic reactions: applications to insect innate immunity. Insect Biochem Mol Biol 2005; 35: 443– 459 [Crossref]
    [Google Scholar]
  10. Zhao P, Li J, Wang Y, Jiang H. Broad-spectrum antimicrobial activity of the reactive compounds generated in vitro by Manduca sexta phenoloxidase. Insect Biochem Mol Biol 2007; 37: 952– 959 [Crossref]
    [Google Scholar]
  11. Zhao P, Lu Z, Strand MR, Jiang H. Antiviral, anti-parasitic, and cytotoxic effects of 5,6-dihydroxyindole (DHI), a reactive compound generated by phenoloxidase during insect immune response. Insect Biochem Mol Biol 2011; 41: 645– 652 [Crossref]
    [Google Scholar]
  12. Charoensapsri W, Amparyup P, Suriyachan C, Tassanakajon A. Melanization reaction products of shrimp display antimicrobial properties against their major bacterial and fungal pathogens. Dev Comp Immunol 2014; 47: 150– 159 [Crossref]
    [Google Scholar]
  13. Eleftherianos I, Boundy S, Joyce SA, Aslam S, Marshall JW et al. An antibiotic produced by an insect pathogenic bacterium suppresses host defenses through phenoloxidase inhibition. Proc Natl Acad Sci USA 2007; 104: 2419– 2424 [Crossref]
    [Google Scholar]
  14. Beck MH, Strand MR. A novel polydnavirus protein inhibits the insect prophenoloxidase activation pathway. Proc Natl Acad Sci USA 2007; 104: 19267– 19272 [Crossref]
    [Google Scholar]
  15. Lu Z, Beck MH, Wang Y, Jiang H, Strand MR. The viral protein Egf1.0 is a dual activity inhibitor of prophenoloxidase-activating proteinases 1 and 3 from Manduca sexta. J Biol Chem 2008; 283: 21325– 21333 [CrossRef] [PubMed]
    [Google Scholar]
  16. Lu Z, Beck MH, Strand MR. Egf1.5 is a second phenoloxidase cascade inhibitor encoded by Microplitis demolitor bracovirus. Insect Biochem Mol Biol 2010; 40: 497– 505 [Crossref]
    [Google Scholar]
  17. Escobedo-Bonilla CM, Alday-Sanz V, Wille M, Sorgeloos P, Pensaert MB et al. Review on the morphology, molecular characterization, morphogenesis and pathogenesis of white spot syndrome virus. J Fish Dis 2008; 31: 1– 18 [Crossref]
    [Google Scholar]
  18. Shekhar MS, Ponniah AG. Recent insights into host-pathogen interaction in white spot syndrome virus infected penaeid shrimp. J Fish Dis 2015; 38: 599– 612 [Crossref]
    [Google Scholar]
  19. Sutthangkul J, Amparyup P, Charoensapsri W, Senapin S, Phiwsaiya K et al. Suppression of shrimp melanization during white spot syndrome virus infection. J Biol Chem 2015; 290: 6470– 6481 [Crossref]
    [Google Scholar]
  20. Ai HS, Huang YC, Li SD, Weng SP, Yu XQ et al. Characterization of a prophenoloxidase from hemocytes of the shrimp Litopenaeus vannamei that is down-regulated by white spot syndrome virus. Fish Shellfish Immunol 2008; 25: 28– 39 [CrossRef] [PubMed]
    [Google Scholar]
  21. Ai HS, Liao JX, Huang XD, Yin ZX, Weng SP et al. A novel prophenoloxidase 2 exists in shrimp hemocytes. Dev Comp Immunol 2009; 33: 59– 68 [CrossRef] [PubMed]
    [Google Scholar]
  22. Yeh SP, Chen YN, Hsieh SL, Cheng W, Liu CH. Immune response of white shrimp, Litopenaeus vannamei, after a concurrent infection with white spot syndrome virus and infectious hypodermal and hematopoietic necrosis virus. Fish Shellfish Immunol 2009; 26: 582– 588 [CrossRef] [PubMed]
    [Google Scholar]
  23. Jiravanichpaisal P, Sricharoen S, Söderhäll I, Söderhäll K. White spot syndrome virus (WSSV) interaction with crayfish haemocytes. Fish Shellfish Immunol 2006; 20: 718– 727 [Crossref]
    [Google Scholar]
  24. Charoensapsri W, Amparyup P, Hirono I, Aoki T, Tassanakajon A. Gene silencing of a prophenoloxidase activating enzyme in the shrimp, Penaeus monodon, increases susceptibility to Vibrio harveyi infection. Dev Comp Immunol 2009; 33: 811– 820 [CrossRef] [PubMed]
    [Google Scholar]
  25. Charoensapsri W, Amparyup P, Hirono I, Aoki T, Tassanakajon A. PmPPAE2, a new class of crustacean prophenoloxidase (proPO)-activating enzyme and its role in PO activation. Dev Comp Immunol 2011; 35: 115– 124 [CrossRef] [PubMed]
    [Google Scholar]
  26. Lan Y, Xu X, Yang F, Zhang X. Transcriptional profile of shrimp white spots syndrome virus (WSSV) genes with DNA microarray. Arch Virol 2006; 151: 1723– 1733 [Crossref]
    [Google Scholar]
  27. Tsai JM, Wang HC, Leu JH, Hsiao HH, Wang AH et al. Genomic and proteomic analysis of thirty-nine structural proteins of shrimp white spot syndrome virus. J Virol 2004; 78: 11360– 11370 [Crossref]
    [Google Scholar]
  28. Roux MM, Pain A, Klimpel KR, Dhar AK. The lipopolysaccharide and β-1,3-glucan binding protein gene is upregulated in white spot virus-infected shrimp (Penaeus stylirostris). J Virol 2002; 76: 7140– 7149 [CrossRef] [PubMed]
    [Google Scholar]
  29. Xie X, Xu L, Yang F. Proteomic analysis of the major envelope and nucleocapsid proteins of white spot syndrome virus. J Virol 2006; 80: 10615– 10623 [Crossref]
    [Google Scholar]
  30. Lin X, Söderhäll I. Crustacean hematopoiesis and the astakine cytokines. Blood 2011; 117: 6417– 6424 [Crossref]
    [Google Scholar]
  31. Lin SW, Chuang YC, Lin YS, Lei HY, Liu HS et al. Dengue virus nonstructural protein NS1 binds to prothrombin/thrombin and inhibits prothrombin activation. J Infect 2012; 64: 325– 334 [Crossref]
    [Google Scholar]
  32. Söderhäll I, Wu C, Novotny M, Lee BL, Söderhäll K. A novel protein acts as a negative regulator of prophenoloxidase activation and melanization in the freshwater crayfish Pacifastacus leniusculus. J Biol Chem 2009; 284: 6301– 6310 [CrossRef] [PubMed]
    [Google Scholar]
  33. Angthong P, Watthanasurorot A, Klinbunga S, Ruangdej U, Söderhäll I et al. Cloning and characterization of a melanization inhibition protein (PmMIP) of the black tiger shrimp, Penaeus monodon. Fish Shellfish Immunol 2010; 29: 464– 468 [CrossRef] [PubMed]
    [Google Scholar]
  34. Christen JM, Hiromasa Y, An C, Kanost MR. Identification of plasma proteinase complexes with serpin-3 in Manduca sexta. Insect Biochem Mol Biol 2012; 42: 946– 955 [Crossref]
    [Google Scholar]
  35. An C, Kanost MR. Manduca sexta serpin-5 regulates prophenoloxidase activation and the Toll signaling pathway by inhibiting hemolymph proteinase HP6. Insect Biochem Mol Biol 2010; 40: 683– 689 [CrossRef] [PubMed]
    [Google Scholar]
  36. Park DS, Shin SW, Hong SD, Park HY. Immunological detection of serpin in the fall webworm, Hyphantria cunea and its inhibitory activity on the prophenoloxidase system. Mol Cells 2000; 10: 186– 192 [CrossRef] [PubMed]
    [Google Scholar]
  37. Xie X, Li H, Xu L, Yang F. A simple and efficient method for purification of intact white spot syndrome virus (WSSV) viral particles. Virus Res 2005; 108: 63– 67 [CrossRef] [PubMed]
    [Google Scholar]
  38. Li S, Zhang X, Sun Z, Li F, Xiang J. Transcriptome analysis on Chinese shrimp Fenneropenaeus chinensis during WSSV acute infection. PLoS One 2013; 8: e58627 [CrossRef] [PubMed]
    [Google Scholar]
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