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Polymorphisms of β-defensin genes in Valle del Belice dairy sheep

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Abstract

The aim of this work was to study β-defensin 1 (SBD1) and β-defensin 2 (SBD2) genes in Valle del Belice dairy sheep in order to identify polymorphisms that can be utilized as markers of the analyzed genes, and search for the functional effects and roles of the identified polymorphisms (variation of the amino acid sequence of the protein and stability of mRNA molecule). The study was conducted on 300 randomly selected animals belonging to four flocks. A total of seven SNPs were identified, two in SBD1 and five in SBD2. The two SNPs identified in SBD2 coding region, at position 1659 and position 1667, were non-synonymous, leading to amino acid changes in the protein product. Nevertheless, the functional effects predicted by the two SNPs demonstrated that amino acid substitutions may not have effect on β-defensin 2 protein function. Moreover, we demonstrated that SBD2 mutant sequence shows changes in mRNA secondary structure. These results suggest that identified SNPs could play a role in the modulation of the immune response.

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Abbreviations

SBD1:

β-Defensin 1 gene

SBD2:

β-Defensin 2 gene

AMPs:

Antimicrobial peptides family

SCC:

Somatic cell count

References

  1. Yount NY, Wang MS, Yuan J, Banaiee N, Oullette AJ, Selsted ME (1995) Rat neutrophil defensins. Precursor structures and expression during neutrophilic myelopoiesis. J Immunol 155:4476–4484

    PubMed  CAS  Google Scholar 

  2. Otvos LJr (2000) Antibacterial peptides isolated from insects. J Pept Sci 6:497–511

    Article  PubMed  CAS  Google Scholar 

  3. Thomma BPHJ, Cammue BPA, Thevissen K (2002) Plant defensins. Planta 216:193–202

    Article  PubMed  CAS  Google Scholar 

  4. Brogden KA, Ackermann M, McCray PBJr, Tack BF (2003) Antimicrobial peptides in animals and their role in host defences. Int J Antimicrob Ag 22:465–478

    Article  CAS  Google Scholar 

  5. Lehrer RI (2004) Primate defensins. Nat Rev Microbiol 2:727–738

    Article  PubMed  CAS  Google Scholar 

  6. Lehrer RI, Ganz T (2002) Defensins of vertebrate animals. Curr Opin Immunol 14:96–102

    Article  PubMed  CAS  Google Scholar 

  7. Selsted ME, Ouellette A (2005) Mammalian defensins in the antimicrobial immune response. Nat Immunol 6:551–557

    Article  PubMed  CAS  Google Scholar 

  8. Kaiser V, Diamond G (2000) Expression of mammalian defensin genes. J Leukoc Biol 68:779–784

    PubMed  CAS  Google Scholar 

  9. Yang D, Biragyn A, Kwak LW, Oppenheim JJ (2002) Mammalian defensins in immunity: more than just microbicidal. Trends Immunol 23:291–296

    Article  PubMed  CAS  Google Scholar 

  10. Sorensen OE, Cowland JB, Theilgaard-Mönch K, Liu L, Ganz T, Borregaard N (2003) Wound healing and expression of antimicrobial peptides/polypeptides in human keratinocytes, a consequence of common growth factors. J Immunol 170:5583–5589

    PubMed  CAS  Google Scholar 

  11. Yang D, Chertov O, Bykovskaia SN, Chen Q, Buffo MJ, Shogan J, Anderson M, Schroder JM, Wang JM, Howard OM, Oppenheim JJ (1999) Beta-defensins: linking innate and adaptive immunity through dendritic and T cell CCR6. Science 286:525–528

    Article  PubMed  CAS  Google Scholar 

  12. Maxwell AI, Morrison GM, Dorin JR (2003) Rapid sequence divergence in mammalian β-defensins by adaptative evolution. Mol Immunol 40:413–421

    Article  PubMed  CAS  Google Scholar 

  13. Patil AA, Cai Y, Sang Y, Blecha F, Zhang G (2005) Cross-species analysis of the mammalian β-defensin gene family: presence of syntenic gene clusters and preferential expression in the male reproductive tract. Physiol Genomics 23:5–17

    Article  PubMed  CAS  Google Scholar 

  14. Diamond G, Kaiser V, Rhodes J, Russell JP, Bevins CL (2000) Transcriptional regulation of β-defensin gene expression in tracheal epithelial cells. Infect Immun 68:113–119

    Article  PubMed  CAS  Google Scholar 

  15. Roosen S, Exner K, Paul S, Schroder JM, Kalm E, Looft C (2004) Bovine β-defensins: identification and characterization of novel bovine β-defensin genes and their expression in mammary gland tissue. Mamm Genome 15:834–842

    Article  PubMed  CAS  Google Scholar 

  16. Selsted ME, Tang YQ, Morris WL, McGuire PA, Novotny MJ, Smith W, Henschen AH, Cullor JS (1993) Purification, primary structures, and antibacterial activities of β-defensins, a new family of antimicrobial peptides from bovine neutrophils. J Biol Chem 268:6641–6648

    PubMed  CAS  Google Scholar 

  17. Yount NY, Yuan J, Tarver A, Castro T, Diamond G, Tran PA, Leavy JN, McCullough C, Cullor JS, Bevins CL, Selsted ME (1999) Cloning and expression of bovine neutrophil β-defensins, biosynthetic profile during neutrophilic maturation and localization of mature peptide to novel cytoplasmic dense granules. J Biol Chem 274:26249–26258

    Article  PubMed  CAS  Google Scholar 

  18. Diamond G, Jones DE, Bevins CL (1993) Airway epithelial cells are the site of expression of a mammalian antimicrobial peptide gene. Proc Natl Acad Sci USA 90:4596–4600

    Article  PubMed  CAS  Google Scholar 

  19. Cormican P, Meade KG, Cahalane S, Narciandi F, Chapwanya A, Lloyd AT, O’Farrelly C (2008) Evolution, expression and effectiveness in a cluster of novel bovine beta-defensins. Immunogenet 60:147–156

    Article  CAS  Google Scholar 

  20. Shi J, Zhang G, Wu H, Ross C, Blecha F, Ganz T (1999) Porcine epithelial β-defensin 1 is expressed in the dorsal tongue at antimicrobial concentrations. Infect Immun 67:3121–3127

    PubMed  CAS  Google Scholar 

  21. Sang Y, Patil AA, Zhang G, Ross CR, Blecha F (2006) Bioinformatic and expression analysis of novel porcine β-defensins. Mamm Genome 17:332–339

    Article  PubMed  CAS  Google Scholar 

  22. Zhao C, Nguyen T, Liu L, Shamova O, Brodgen K, Lehrer RI (1999) Differential expression of caprine β defensins in digestive and respiratory tissues. Infect Immun 67:6221–6224

    PubMed  CAS  Google Scholar 

  23. Sharma A, Dev K, Kumar A (2006) Cloning and characterization of goat lingual antimicrobial peptide (Unpublished) (GenBank DQ836129)

  24. Huttner KM, Lambeth MR, Burkin HR, Burkin DJ, Broad TE (1998) Localization and genomic organization of sheep antimicrobial peptide genes. Gene 206:85–91

    Article  PubMed  CAS  Google Scholar 

  25. Luenser K, Fickel J, Ludwig A (2005) Evolution of caprine and ovine β-defensin genes. Immunogenet 57:487–498

    Article  CAS  Google Scholar 

  26. Guimarães RL, Segat L, Rocha CRC, Brandão LAC, Zanin V, Araujo J, Naslavsky MS, de Lima Filho JL, Crovella S (2009) Functional polymorphisms of DEFB1 gene in type 1 diabetes Brazilian children. Autoimmun 42:406–413

    Article  Google Scholar 

  27. Ricci E, Malacrida S, Zanchetta M, Montagna M, Giaquinto C, De Rossi A (2009) Role of beta-defensin-1 polymorphisms in mother-to-child transmission of HIV-1. J Acquir Immune Defic Syndr 51:13–19

    Article  PubMed  CAS  Google Scholar 

  28. Ryniewicz Z, Zwierzchowski L, Bagnicka E, Flisikowski K, Maj A, Krzyżewski J, Strzałkowska N (2003) Association of the polymorphism at defensin gene loci with dairy production traits and milk somatic cell count in Black-and-White cows. Anim Sci Pap Rep 21:209–222

    CAS  Google Scholar 

  29. Wojdak-Maksymiec K, Kmieć M, Żukiewicz A (2006) Associations between defensin polymorphism and somatic cell count in milk and milk utility traits in Jersey dairy cows. J Vet Med A 53:495–500

    Article  CAS  Google Scholar 

  30. Bagnicka E, Strzalkowska N, Flisikowski K, Szreder T, Jozwik A, Prusak B, Krzyzewski J, Zwierzchowski L (2007) The polymorphism in the beta4-defensin gene and its association with production and somatic cell count in Holstein-Friesian cows. J Anim Breed Genet 124:150–156

    Article  PubMed  CAS  Google Scholar 

  31. Miller SA, Dykes DD, Polesky HF (1988) A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 16:1215

    Article  PubMed  CAS  Google Scholar 

  32. 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 22:4673–4680

    Article  PubMed  CAS  Google Scholar 

  33. Rousset F (2008) Genepop’007: a complete re-implementation of the Genepop software for Windows and Linux. Mol Ecol Resour 8:103–106

    Article  PubMed  Google Scholar 

  34. Thomas PD, Campbell MJ, Kejariwal A, Mi H, Karlak B, Daverman R, Diemer K, Muruganujan A, Narechania A (2003) PANTHER: a library of protein families and subfamilies indexed by function. Genome Res 13:2129–2141

    Article  PubMed  CAS  Google Scholar 

  35. Ng PC, Henikoff S (2003) SIFT: predicting amino acid changes that affect protein function. Nucleic Acids Res 31:3812–3814

    Article  PubMed  CAS  Google Scholar 

  36. Mathews DH, Disney MD, Childs JL, Schroeder SJ, Zuker M, Turner DH (2004) Incorporating chemical modification constraints into a dynamic programming algorithm for prediction of RNA secondary structure. Proc Natl Acad Sci USA 101:7287–7292

    Article  PubMed  CAS  Google Scholar 

  37. Bowdish DME, Davidson DJ, Scott MG, Hancock REW (2005) Immunomodulatory activities of small host defense peptides. Antimicrob Agents Chemother 49:1727–1732

    Article  PubMed  CAS  Google Scholar 

  38. Pazgier M, Prah A, Hoover DM, Lubkowski J (2006) Studies of the biological properties of human β-defensin 1. J Biol Chem 282:1819–1829

    Article  PubMed  Google Scholar 

  39. Zou G, de Leeuw E, Li C, Pazgier M, Li C, Zeng P, Wei-Yue L, Lubkowski J, Lu W (2007) Toward understanding the cationicity of defensins arg and lys versus their noncoded analogs. J Biol Chem 282:19653–19665

    Article  PubMed  CAS  Google Scholar 

  40. Kalus AA, Fredericks LP, Hacker BM, Dommisch H, Presland RB, Kimball JR, Dale BA (2009) Association of a genetic polymorphism (−44 C/G SNP) in the human DEFB1 gene with expression and inducibility of multiple β-defensins in gingival keratinocytes. BMC Oral Health. doi:10.1186/1472-6831-9-21

  41. Hafez EE, Abdel-Rahman SM, El-Sohaimy SA (2008) Accompaniment of the polymorphism at defensin gene loci with milk productivity in holstein friesian and egyptian cows. Biotechnol Anim Husb 24:9–21

    Article  Google Scholar 

  42. Bergonier D, de Cremoux R, Rupp R, Lagriffoul G, Berthelot X (2003) Mastitis of dairy small ruminants. Vet Res 34:689–716

    Article  PubMed  Google Scholar 

  43. Cartegni L, Shern LC, Krainer AR (2002) Listening to silence and understanding nonsense: exonic mutations that affect splicing. Nat Rev 3:285–298

    Article  CAS  Google Scholar 

  44. Chamary JV, Parmley JL, Hurst LD (2006) Hearing silence: non-neutral evolution at synonymous sites in mammals. Nat Rev Genet 7:98–108

    Article  PubMed  CAS  Google Scholar 

  45. Fujii G, Selsted ME, Eisenberg D (1993) Defensins promote fusion and lysis of negatively charged membranes. Protein Sci 2:1301–1312

    Article  PubMed  CAS  Google Scholar 

  46. Semple CAM, Rolfe M, Dorin JR (2003) Duplication and selection in the evolution of primate β-defensin genes. Genome Biol 4:R31.1-.11

    Google Scholar 

  47. Raj PA, Antonyraj KJ, Karunakaran T (2000) Large-scale synthesis and functional elements for the antimicrobial activity of defensins. Biochem J 347:633–641

    Article  PubMed  CAS  Google Scholar 

  48. Boniotto M, Antcheva N, Zelezetsky I, Tossi A, Palumbo V, Verga Falzacappa MV, Sgubin S, Braida L, Amoroso A, Crovella S (2003) A study of host defence peptide β-defensin 3 in primates. Biochem J 374:707–714

    Article  PubMed  CAS  Google Scholar 

  49. Klüver E, Schulz-Maronde S, Scheid S, Meyer B, Forssmann WG, Adermann K (2005) Structure-activity relation of human β-defensin 3: influence of disulfide bonds and cysteine substitution on antimicrobial activity and cytotoxicity. Biochemistry 44:9804–9816

    Article  PubMed  Google Scholar 

  50. Klüver E, Adermann K, Schulz A (2006) Synthesis and structure-activity relationship of β-defensins, multi-functional peptides of the immune system. J Pept Sci 12:243–257

    Article  PubMed  Google Scholar 

  51. van der Velden AW, Thomas AA (1999) The role of 5′ untraslated region of an mRNA in translation regulating during development. Int J Biochem Cell B 31:87–106

    Article  Google Scholar 

  52. Jansen RP (2001) mRNA localization: message on the move. Nat Rev Mol Cell Biol 2:247–256

    Article  PubMed  CAS  Google Scholar 

  53. Bashirullah A, Cooperstock RL, Lipshitz HD (2001) Spatial and temporal control of RNA stability. Proc Natl Acad Sci USA 98:7025–7028

    Article  PubMed  CAS  Google Scholar 

  54. Conne B, Stuzt A, Vassalli JD (2000) The 3′ untraslated region of messenger RNA: a molecular ‘hotspots’ for pathology? Nat Med 6:637–641

    Article  PubMed  CAS  Google Scholar 

  55. Puga I, Lainez B, Fernandez-Real JM, Buxade M, Broch M, Vendrell J, Espel E (2005) A polymorphism in the 3′ untraslated region of the gene for tumor necrosis factor receptor 2 modulated reporter gene expression. Endocrinology 146:2210–2220

    Article  PubMed  CAS  Google Scholar 

  56. Wang J, Pitarque M, Ingelman-Sundberg M (2006) 3′-UTR polymorphism in the human CYP2A6 gene affect mRNA stability and enzyme expression. Biochem Biophys Res Commun 340:491–497

    Article  PubMed  CAS  Google Scholar 

  57. Prado-Montes de Oca E, Velarde-Felix JS, Rios-Tostado JJ, Picos-Cardenas VJ, Figuera LE (2009) SNP 668C (−44) alters a NF-kappaB1 putative binding site in non-coding strand of human beta-defensin 1 (DEFB1) and is associated with lepromatous leprosy. Infect Genet Evol 9:617–625

    Article  PubMed  CAS  Google Scholar 

  58. Schaefer AS, Richter GM, Nothnagel M, Laine ML, Rühling A, Schäfer C, Cordes N, Noack B, Folwaczny M, Glas J, Dörfer C, Dommisch H, Groessner-Schreiber B, Jepsen S, Loos BG, Schreiber S (2010) A 3′ UTR transition within DEFB1 is associated with chronic and aggressive periodontitis. Genes Immun 11:45–54

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The authors would like to acknowledge the Ministero delle Politiche Agricole Alimentari e Forestali (MiPAAF) (D.M. 302/7303/05) and the Ministero dell’Istruzione, dell’Università e della Ricerca (project #2007898KYN, PRIN 2007) for financial support for this research.

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Authors and Affiliations

  1. Dipartimento DEMETRA-Sezione Produzioni Animali, Università di Palermo, Viale delle Scienze Parco d’Orleans, 90128, Palermo, Italy

    Giuseppina Monteleone, Davide Calascibetta, Paola Galluzzo, Valentina Riggio & Baldassare Portolano

  2. Consorzio Regionale di Ricerca Bioevoluzione Sicilia, c/o Dipartimento DEMETRA-Sezione Produzioni Animali, Università di Palermo, Viale delle Scienze Parco d’Orleans, 90128, Palermo, Italy

    Davide Calascibetta, Mariangela Scaturro & Marisa Palmeri

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  1. Giuseppina Monteleone
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  2. Davide Calascibetta
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  3. Mariangela Scaturro
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  4. Paola Galluzzo
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Correspondence to Giuseppina Monteleone.

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Giuseppina Monteleone and Davide Calascibetta equally contributed to this work.

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Monteleone, G., Calascibetta, D., Scaturro, M. et al. Polymorphisms of β-defensin genes in Valle del Belice dairy sheep. Mol Biol Rep 38, 5405–5412 (2011). https://doi.org/10.1007/s11033-011-0694-5

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  • DOI: https://doi.org/10.1007/s11033-011-0694-5

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