Fisheries and Aquatic Sciences

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Isolation of Novel Hepcidin Isoforms from the Rockbream Oplegnathus fasciatus (Perciformes)

  • Lee, Sang-Yoon (Department of Marine Bio-Materials and Aquaculture, Pukyong National University) ;
  • Nam, Yoon-Kwon (Department of Marine Bio-Materials and Aquaculture, Pukyong National University)
  • Received : 2011.02.07
  • Accepted : 2011.03.03
  • Published : 2011.03.31
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Abstract

Three novel hepcidin isoforms were isolated and characterized from the perciform fish species Oplegnathus fasciatus. These hepcidin isoforms (designated rbhepc5, rbhepc6 and rbhepc7) were found to share a conserved, tripartite gene structure and a considerable sequence homology one another. A comparison of their mature peptide sequences with those of other perciform hepcidin orthologs indicated that these three hepcidin isoforms as well as four other isoforms previously identified in this species, appear to belong to the HAMP2 group of hepcidin genes. Analysis of the 5'-upstream sequences showed that the proximal non-coding regions of rbhepc5~7 do not possess canonical TATA signals; instead, they harbor several binding motifs for transcription factors involved in immune modulation. Reverse transcriptase-PCR analysis demonstrated that the rbhepc5~7 are expressed predominantly in the liver, and that the transcription of rbhepc5~7 is rapidly induced in the liver, but not in other tissues, by experimental challenge with any of three different bacterial species. However, transcription of rbhepc6 appeared to be negligible under both basal and stimulated conditions, as judged by the redundancy count of randomly chosen reverse transcriptase-PCR clones.

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References

  1. Anderson KV. 2000. Toll signaling pathways in the innate immune response. Curr Opin Immunol 12, 13-19. https://doi.org/10.1016/S0952-7915(99)00045-X
  2. Atanasiu V, Manolescu B and Stoian I. 2006. Hepcidin - central regulator of iron metabolism. Eur J Haematol 78, 1-10.
  3. Bao B, Peatman E, Li P, He C and Liu Z. 2005. Catfish hepcidin gene is expressed in a wide range of tissues and exhibits tissue-specific upregulation after bacterial infection. Dev Comp Immunol 29, 939-950. https://doi.org/10.1016/j.dci.2005.03.006
  4. Cho YS, Lee SY, Kim KH, Kim SK, Kim DS and Nam YK. 2009. Gene structure and differential modulation of multiple rockbream (Oplegnathus fasciatus) hepcidin isoforms resulting from different biological stimulations. Dev Comp Immunol 33, 46-58. https://doi.org/10.1016/j.dci.2008.07.009
  5. Choi SK, Kwon SR, Nam YK, Kim SK and Kim KH. 2006. Organ distribution of red sea bream iridovirus (RSIV) DNA in asymptomatic yearling and fingerling rock bream (Oplegnathus fasciatus) and effects of water temperature on transition of RSIV into acute phase. Aquaculture 256, 23-26. https://doi.org/10.1016/j.aquaculture.2006.01.026
  6. Courselaud B, Pigeon C, Inoue Y, Inoue J, Gonzalez FJ, Leroyer P, Gilot D, Boudjema K, Guguen-Guillouzo C, Brissot P, Loreal O and Ilyin G. 2002. $C/EBP{\alpha}$ regulates hepatic transcription of hepcidin, an antimicrobial peptide and regulator of iron metabolism. J Biol Chem 277, 41163-41170. https://doi.org/10.1074/jbc.M202653200
  7. Cuesta A, Meseguer J and Esteban MA. 2008. The antimicrobial peptide hepcidin exerts an important role in the innate immunity against bacteria in thebony fish gilthead seabream. Mol Immunol 45, 2333-2342. https://doi.org/10.1016/j.molimm.2007.11.007
  8. Douglas SE, Gallant JW, Liebscher RS, Dacanay A and Tsoi SC. 2003. Identification and expression analysis of hepcidin-like antimicrobial peptides in bony fish. Dev Comp Immunol 27, 589-601. https://doi.org/10.1016/S0145-305X(03)00036-3
  9. Hilton KB and Lambert LA. 2008. Molecular evolution and characterization of hepcidin gene products in vertebrates. Gene 415, 40-48. https://doi.org/10.1016/j.gene.2008.02.016
  10. Hsieh J, Pan C and Chen J. 2010. Tilapia hepcidin (TH) 2-3 as a transgene in transgenic fish enhances resistance to vibrio vulnificus infection and causes variations in immune-related genes after infection by different bacterial species. Fish Shellfish Immunol 29, 430-439. https://doi.org/10.1016/j.fsi.2010.05.001
  11. Huang P, Chen J and Kuo C. 2007. Three different hepcidins from tilapia, Oreochromis mossambicus: Analysis of their expressions and biological functions. Mol Immunol 44, 1922-1934. https://doi.org/10.1016/j.molimm.2006.09.031
  12. Lauth X, Babon JJ, Stannard JA, Singh S, Nizet V, Carlberg JM, Ostland VE, Pennington MW, Norton RS and Westerman ME. 2005. Bass hepcidin synthesis, solution structure, antimicrobial activities and synergism, and in vivo hepatic response to bacterial infections. J Biol Chem 280, 9272-9282. https://doi.org/10.1074/jbc.M411154200
  13. Lee SY and Nam YK. 2009. Stimulated mRNA expression of the second glyceraldehyde 3-phosphate dehydrogenase in the barred knifejaw Oplegnathus fasciatus spleen during bacterial and viral injection. J Aquaculture 22, 51-55.
  14. Nemeth E, Preza GC, Jung C, Kaplan J, Waring AJ and Ganz T. 2006. The N-terminus of hepcidin is essential for its interaction with ferroportin: Structure-function study. Blood 107, 328-333. https://doi.org/10.1182/blood-2005-05-2049
  15. Nemeth E, Tuttle MS, Powelson J, Vaughn MB, Donovan A, Ward DM, Ganz T and Kaplan J. 2004. Hepcidin regulates cellular iron efflux by binding to ferroportinand inducing its internalization. Science 306, 2090-2093. https://doi.org/10.1126/science.1104742
  16. Padhi A and Verghese B. 2007. Evidence for positive Darwinian selection on the hepcidin gene of Perciform and Pleuronectiform fishes. Mol Divers 11, 119-130. https://doi.org/10.1007/s11030-007-9066-4
  17. Park CH, Valore EV, Waring AJ and Ganz T. 2001. Hepcidin, a urinary antimicrobial peptide synthesized in the liver. J Biol Chem 276, 7806-7810. https://doi.org/10.1074/jbc.M008922200
  18. Ren H, Wang K, Zhou H and Yang M. 2006. Cloning and organisation analysis of a hepcidin-like gene and cDNA from Japan sea bass, Lateolabrax japonicus. Fish Shellfish Immunol 21, 221-227. https://doi.org/10.1016/j.fsi.2005.10.011
  19. Rodrigues PNS, Vazquez-Dorado S, Neves JV and Wilson JM. 2006. Dual function of fish hepcidin: Response to experimental iron overload and bacterial infection in sea bass (Dicentrarchus labrax). Dev Comp Immunol 30, 1156-1167. https://doi.org/10.1016/j.dci.2006.02.005
  20. Sambrook J and Russell DW. 2001. Molecular Cloning: A laboratory manual (3rd ed). Cold Spring Harbor Laboratory Press, New York.
  21. Shi J and Camus AC. 2006. Hepcidins in amphibians and fishes: Antimicrobial peptides or iron-regulatory hormones? Dev Comp Immunol 30, 746-755. https://doi.org/10.1016/j.dci.2005.10.009
  22. Shike H, Shimizu C, Lauth X and Burns JC. 2004. Organization and expression analysis of the zebrafish hepcidin gene, an antimicrobial peptide gene conserved among vertebrates. Dev Comp Immunol 28, 747-754. https://doi.org/10.1016/j.dci.2003.11.009
  23. Shike H, Lauth X, Westerman ME, Ostland VE, Carlberg JM, Van Olst JC, Shimizu C, Bulet P and Burns JC. 2002. Bass hepcidin is a novel antimicrobial peptide induced by bacterial challenge. Eur J Biochem 269, 2232-2237. https://doi.org/10.1046/j.1432-1033.2002.02881.x
  24. Tennessen JA. 2005. Molecular evolution of animal antimicrobial peptides: widespread moderate positive selection. J Evol Biol 18, 1387-1394. https://doi.org/10.1111/j.1420-9101.2005.00925.x
  25. Tossi A, Sandri L and Giangaspero A. 2000. Amphipathic, $\alpha$-helical antimicrobial peptides. Biopolymers 55, 4-30. https://doi.org/10.1002/1097-0282(2000)55:1<4::AID-BIP30>3.0.CO;2-M
  26. Truksa J, Lee P and Beutler E. 2007. The role of STAT, AP-1, E-box and TIEG motifs in the regulation of hepcidin by IL-6 and BMP-9: Lessons from human HAMP and murine Hamp1 and Hamp2 gene promoters. Blood Cell Mol Dis 39, 255-262. https://doi.org/10.1016/j.bcmd.2007.06.014
  27. Valore EV and Ganz T. 2008. Posttranslational processing of hepcidin in human hepatocytes is mediated by the prohormone convertase furin. Blood Cell Mol Dis 40, 132-138. https://doi.org/10.1016/j.bcmd.2007.07.009
  28. Wrighting DM and Andrews NC. 2006. Interleukin-6 induces hepcidin expression through STAT3. Blood 108, 3204-3209. https://doi.org/10.1182/blood-2006-06-027631
  29. Xu Q, Cheng CC, Hu P, Ye H, Chen Z, Cao L, Chen L, Shen Y and Chen L. 2008. Adaptive evolution of hepcidin genes in Antarctic notothenioid fishes. Mol Biol Evol 25, 1099-1112. https://doi.org/10.1093/molbev/msn056
  30. Yang M, Wang K, Chen J, Qu H and Li S. 2007. Genomic organization and tissue-specific expression analysis of hepcidin-like genes from black porgy (Acanthopagrus schlegelii B). Fish Shellfish Immunol 23, 1060-1071. https://doi.org/10.1016/j.fsi.2007.04.011