, 59:233 | Cite as

A gene family of putative immune recognition molecules in the hydroid Hydractinia

  • Ryan S. Schwarz
  • Linda Hodes-Villamar
  • Kelly A. Fitzpatrick
  • Matthew G. Fain
  • Austin L. Hughes
  • Luis F. Cadavid
Original Paper


Animal taxa display a wide array of immune-type receptors that differ in their specificities, diversity, and mode of evolution. These molecules ensure effective recognition of potential pathogens for subsequent neutralization and clearance. We have characterized a family of putative immune recognition molecules in the colonial hydroid Hydractinia symbiolongicarpus. A complementary DNA fragment with high similarity to the sea urchin l-rhamnose-binding lectin was isolated and used to screen 9.5 genome equivalents of a H. symbiolongicarpus bacterial artificial chromosome library. One of the resulting 19 positive clones was sequenced and revealed the presence of a 5,111-bp gene organized in 13 exons and 12 introns. The gene was predicted to encode a 726-amino acid secreted modular protein composed of a signal peptide, an anonymous serine-rich domain, eight thrombospondin type 1 repeats, and a l-rhamnose-binding lectin domain. The molecule was thus termed Rhamnospondin (Rsp). Southern hybridization and sequence analyses indicated the presence of a second Rsp gene. The cDNA from both Rsp genes was sequenced in 18 individuals, revealing high levels of genetic polymorphism. Nucleotide substitutions were distributed throughout the molecule and showed a significantly higher number of synonymous substitutions per synonymous sites than its nonsynonymous counterparts. Whole-mount in situ hybridization and semi-quantitative reverse transcription polymerase chain reaction of microorganism-challenged colonies indicated that Rsp molecules were specifically and constitutively expressed in the hypostome of gastrozooids’ mouth. Thus, the combination of (1) comparative analysis on domain composition and function, (2) polymorphism, and (3) expression patterns, suggest that Rsp genes encode a family of putative immune recognition receptors, which may act by binding microorganisms invading the colony through the polyp’s mouth.


Hydractinia Invertebrate immunity TSR superfamily Rhamnose-binding lectin Polymorphism 



This work was supported by an NSF award to LFC (IBN0315968). We acknowledge technical support from the University of New Mexico’s Molecular Biology Facility which is supported by NIH grant number 1P20RR18754 from the Institute Development Award (IDeA) Program of the National Center for Research Resources.


  1. Adams JC, Tucker RP (2000) The thrombospondin type 1 repeat (TSR) superfamily: diverse proteins with related roles in neural development. Dev Dyn 218:280–299PubMedCrossRefGoogle Scholar
  2. Ballard WW (1942) The mechanisms of synchronous spawning in Hydractinia and Perannia. Biol Bull 82:329–339CrossRefGoogle Scholar
  3. Barondes SH, Cooper DNW, Gitt MA, Leffler H (1994) Galectins, structure and function of a large family of animal lectins that decipher glycocodes. J Biol Chem 269:20807–20810PubMedGoogle Scholar
  4. Berking S (1991) Control of metamorphosis and pattern formation in Hydractinia (Hydrozoa, Cnidaria). BioEssays 13:323–329CrossRefGoogle Scholar
  5. Bisset KA, Vickerstaff J (1967) Significance of the characteristic chemical pattern of Gram positive and Gram negative bacterial cell walls. Nature 215:1286–1287PubMedCrossRefGoogle Scholar
  6. Booy A, Haddow JD, Olafson RW (2005) Isolation of the salmonid rhamnose-binding lectin STL2 from spores of the microsporidian fish parasite Loma salmonae. J Fish Dis 28:455–462PubMedCrossRefGoogle Scholar
  7. Buss LW, Yund PO (1989) A sibling group of Hydractinia in the northeastern United States. J Mar Biol Assoc UK 69:875–895CrossRefGoogle Scholar
  8. Cadavid LF, Powell AE, Nicotra ML, Moreno M, Buss LW (2004) An invertebrate histocompatibility complex. Genetics 167:357–365PubMedCrossRefGoogle Scholar
  9. Cannon JP, Haire RN, Litman GW (2002) Identification of diversified genes that contain immunoglobulin-like variable regions in a protochordate. Nat Immunol 3:1200–1207PubMedCrossRefGoogle Scholar
  10. Cannon JP, Haire RN, Schnitker N, Mueller MG, Litman GW (2004) Individual protochordates posses unique immune-type receptor repertoires. Curr Biol 14:R465–R466PubMedCrossRefGoogle Scholar
  11. Dawson DW, Pearce SFA, Zhong R, Silverstain RL, Frazier WA, Bouck NP (1997) CD36 mediates the in vitro inhibitory effects of thrombospondin-1 on endothelial cells. J Cell Biol 138:707–717PubMedCrossRefGoogle Scholar
  12. Day AJ (1994) The C-type carbohydrate recognition domain (CRD) superfamily. Biochem Soc Trans 22:83–88PubMedGoogle Scholar
  13. De Tomaso AW, Nyholm SV, Palmeri KJ, Ishizuka KJ, Ludington WB, Mitchel K, Weissman IL (2005) Isolation and characterization of a protochordate histocompatibility locus. Nature 438:454–459PubMedCrossRefGoogle Scholar
  14. Frank U, Leitz T, Muller WA (2001) The hydroid Hydractinia: a versatile, informative cnidarian representative. BioEssays 23:963–971PubMedCrossRefGoogle Scholar
  15. Goundis D, Reid KB (1988) Properdin, the terminal complement complements, thrombospondin and the circumsporozoite protein of malaria parasites contain similar sequence motifs. Nature 335:82–85PubMedCrossRefGoogle Scholar
  16. He Y, Li H, Zhang J, Hsu C, Lin E, Zhang N, Guo J, Forbush KA, Bevan MJ (2004) The extracellular matrix protein mindin is a pattern-recognition molecule for microbial pathogens. Nat Immunol 5:88–97PubMedCrossRefGoogle Scholar
  17. Hosono M, Ishikawa K, Mineki R, Murayama K, Numata C, Ogawa Y, Takayanagi Y, Nitta K (1999) Tandem repeat structure of rhamnose-binding lectin from catfish (Silurus asotus) eggs. Biochem Biophys Acta 1472:668–675PubMedGoogle Scholar
  18. Janeway CA, Medzhitov R (2002) Innate immune recognition. Annu Rev Immunol 20:197–216PubMedCrossRefGoogle Scholar
  19. Kitchens RL, Thompson PA (2005) Modulatory effects of sCD14 and LBP on LPS-host cell interactions. J Endotoxin Res 11:225–229PubMedCrossRefGoogle Scholar
  20. Kornfeld S (1992) Structure and function of the mannose 6-phosphate/insulin-like growth factor II receptors. Annu Rev Biochem 61:307–330PubMedCrossRefGoogle Scholar
  21. Kudo S, Inoue M (1986) A bactericidal effect of fertilization envelope extract from fish egg. Zool Science 3:323–329Google Scholar
  22. Kumar S, Tamura K, Nei M (2004) MEGA3: Integrated software for molecular evolutionary genetic analysis and sequence alignment. Brief Bioinfor 5:150–163CrossRefGoogle Scholar
  23. Le Y, Lee SH, Kon OL, Lu J (1998) Human L-ficolin: Plasma levels, sugar specificity, and assignment of its lectin activity to the fibrinogen-like (FBG) domain. FEBS Lett 425:367–370PubMedCrossRefGoogle Scholar
  24. Litman GW, Cannon JP, Dishaw LJ (2005) Reconstructing immune phylogeny: new perspectives. Nat Rev Immunol 5:866–879PubMedCrossRefGoogle Scholar
  25. Margalit H, Fischer N, Ben-Sasson SA (1993) Comparative analysis of structurally defined heparin binding sequences reveals a distinct spatial distribution of basic residues. J Biol Chem 268:19228–19231PubMedGoogle Scholar
  26. Murphy-Ullrich JE, Poczatek M (2000) Activation of latent TGF-β by thrombospondin-1: mechanisms and physiology. Cytokine Growth Factor Rev 11:59–69PubMedCrossRefGoogle Scholar
  27. Nair SV, Del Valle H, Gross PS, Terwilliger DP, Smith LC (2005) Macroarray analysis of coelomocyte gene expression in response to LPS in the sea urchin. Identification of unexpected immune diversity in an invertebrate. Physiol Genomics 22:33–47PubMedCrossRefGoogle Scholar
  28. Nei M, Gojobori T (1986) Simple methods for estimating the numbers of synonymous and nonsynonymous substitutions. Mol Biol Evol 3:418–426PubMedGoogle Scholar
  29. Nei M, Kumar S (2000) Molecular evolution and phylogenetics. Oxford University Press, OxfordGoogle Scholar
  30. Okamoto M, Tsutsui S, Tasumi S, Suetake H, Kikuchi K, Suzuki M (2005) Tandem repeat l-rhamnose-binding lectin from the skin mucus of ponyfish, Leiognathus nuchalis. Biochem Biophys Res Commun 333:463–469PubMedCrossRefGoogle Scholar
  31. Ozeki Y, Matsui T, Suzuki M, Titani K (1991) Amino acid sequence and molecular charcterization of a D-galactose-specific lectin purified from sea urchin (Anthocidaris crassispina) eggs. Biochemistry 30:2391–2394PubMedCrossRefGoogle Scholar
  32. Ozinsky A, Underhill DM, Fontenot JD, Hajjar AM, Smith KD, Wilson CB, Schroeder L, Aderem A (2000) The repertoire for pattern recognition of pathogens by the innate immune system is defined by cooperation between Toll-like receptors. Proc Natl Acad Sci U S A 97:13766–13771PubMedCrossRefGoogle Scholar
  33. Powell LD, Varki A (1995) I-type lectins. J Biol Chem 270:14243–14246PubMedCrossRefGoogle Scholar
  34. Prakobphol A, Linzer R (1980) Purification and characterization of a rhamnose-containing cell wall antigen from Streptococcus mutans B13 (serotype d). Infect Immun 27:150–157PubMedGoogle Scholar
  35. Quesenberry MS, Ahmed H, Elola M, O’leary N, Vasta GR (2003) Diverse lectine repertoires in tunicates mediate broad recognition and effector innate immune response. Integr Comp Biol 43:323–330CrossRefGoogle Scholar
  36. Rozas J, Sanchez-DelBarrio JC, Messeguer X, Rozas R (2003) DnaSP, DNA sequence polymorphism analyses by the coalescent and other methods. Bioinformatics 19:2496–2497PubMedCrossRefGoogle Scholar
  37. Shafikhani S (2002) Factors affecting PCR-mediated recombination. Environ Microbiol 4:482–486PubMedCrossRefGoogle Scholar
  38. Shiina N, Tateno H, Ogawa T, Muramoto K, Saneyoshi M, Kamiya H (2002) Isolation and characterization of l-rhamnose-binding lectins from chum salmon (Oncorhynchus keta) eggs. Fish Sci 68:1352–1366CrossRefGoogle Scholar
  39. Silverstein RL (2002) The face of TSR revealed: an extracellular signaling domain is exposed. J Cell Biol 159:203–205PubMedCrossRefGoogle Scholar
  40. Steel DM, Whitehead AS (1994) The major acute phase reactant: C-reactive protein, serum amyloid P component and serum amyloid A protein. Immunol Today 15:81–88PubMedCrossRefGoogle Scholar
  41. Suzuki Y, Tasumi S, Tsutsui S, Okamoto M, Suetake H (2003) Molecular diversity of skin mucus lectins in fish. Comp Biochem Physiol 136B:723–730Google Scholar
  42. Tajima F (1989) Statistical method for testing the neutral mutation hypothesis by DNA polymorphism. Genetics 123:585–595PubMedGoogle Scholar
  43. Tan K, Duquette M, Liu J, Dong Y, Zhang R, Joachimiak A, Lawler J, Wang J (2002) Crystal structure of the TSP-1 type 1 repeats: a novel layered fold and its biological implication. J Cell Biol 159:373–382PubMedCrossRefGoogle Scholar
  44. Tateno H, Ogawa T, Muramoto K, Kamiya H, Hirai T, Saneyoshi M (2001) A novel rhamnose-binding lectin family from eggs of steelhead trout (Oncorynchus mykiss) with different structure and tissue distribution. Biosci Biotechnol Biochem 65:1328–1338PubMedCrossRefGoogle Scholar
  45. Tateno H, Ogawa T, Kuramoto K, Kamiya H, Saneyoshi M (2002a) Rhamnose-binding lectin from steelehad trout (Oncorhynchus mykiss) eggs recognize bacterial lipopolysaccharides and lipoteichoic acid. Biosci Biotechnol Biochem 66:604–612PubMedCrossRefGoogle Scholar
  46. Tateno H, Ogawa T, Muramoto K, Kamiya H, Saneyoshi M (2002b) Distribution and molecular evolution of rhamnose-binding lectins in Salmonidae: Isolation and characterization of two lectins from white-spotted charr (Salvelinus leucomaenis) eggs. Biosci Biotechnol Biochem 66:1356–1365PubMedCrossRefGoogle Scholar
  47. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680PubMedCrossRefGoogle Scholar
  48. Tucker RP (2004) The thrombospondin type 1 repeat superfamily. Int J Biochem Cell Biol 36:969–974PubMedCrossRefGoogle Scholar
  49. Wang JX, Zhao XF (2004) Progress in pattern recognition receptors of innate immunity in invertebrates. Prog Biochem Biophys 31:112–117Google Scholar
  50. Watson FL, Puttmann-Holgado R, Thomas F, Lamar DL, Hughes M, Kondo M, Rebel VI, Schmucker D (2005) Extensive diversity of Ig-superfamily proteins in the immune system of insects. Science 309:1874–1878PubMedCrossRefGoogle Scholar
  51. Zaphiropoulos PG (1998) Non-homologous recombination mediated by Thermus aquaticus DNA polymerase I. Evidence supporting a copy choice mechanisms. Nucleic Acids Res 26:2843–2848PubMedCrossRefGoogle Scholar
  52. Zhang SM, Adema CM, Kepler TB, Loker ES (2004) Diversification of Ig superfamily genes in an invertebrate. Science 305:251–254PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Ryan S. Schwarz
    • 1
  • Linda Hodes-Villamar
    • 1
  • Kelly A. Fitzpatrick
    • 1
  • Matthew G. Fain
    • 1
  • Austin L. Hughes
    • 2
  • Luis F. Cadavid
    • 1
    • 3
  1. 1.Department of BiologyThe University of New MexicoAlbuquerqueUSA
  2. 2.Department of Biological SciencesUniversity of South CarolinaColumbiaUSA
  3. 3.Departamento de Biología and Instituto de GenéticaUniversidad Nacional de ColombiaBogota, DCColombia

Personalised recommendations