Skip to main content
Log in

Bioinformatic and expression analysis of novel porcine β-defensins

  • Published:
Mammalian Genome Aims and scope Submit manuscript

Abstract

β-Defensins are a major group of mammalian antimicrobial peptides. Although more than 30 β-defensins have been identified in humans, only one porcine β-defensin has been reported. In this article we report the identification and initial characterization of 11 novel porcine β-defensins (pBD). Using bioinformatic approaches, we screened 287,821 porcine expressed sequence tags for similarity of their predicted peptides to known human β-defensins and identified full-length or partial sequences for the 11 novel pBDs. Similar to the previously identified pBD1, all of these peptides have a consensus β-defensin motif. A differential expression pattern for these newly identified genes was found. For example, unlike most β-defensins, pBD2 and pBD3 were expressed in bone marrow and in other lymphoid tissues including thymus, spleen, lymph nodes, duodenum, and liver. Including pBD2 and pBD3, six porcine β-defensins were expressed in lung and skin. Several newly identified porcine β-defensins, including pBD123, pBD125, and pBD129, were expressed in male reproductive tissues, including lobuli testis and some segments of the epididymis. Phylogenetic analysis indicates that in most cases the evolutionary relationship between individual porcine β-defensins and their human orthologs is closer than the relationship among β-defensins in the same species. These findings establish the existence of multiple porcine β-defensins and suggest that the pig may be an ideal model for the characterization of β-defensin diversity and function.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

DEFB =:

human β-defensin;

EST =:

expressed sequence tag;

GAPDH =:

glyceraldehyde 3-phosphate dehydrogenase;

pBD =:

porcine β-defensin;

RT-PCR =:

reverse-transcriptase polymerase chain reaction

References

  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215: 403–410

    Article  CAS  PubMed  Google Scholar 

  • Boman HG (1995) Peptide antibiotics and their role in innate immunity. Annu Rev Immunol 13: 61–92

    Article  CAS  PubMed  Google Scholar 

  • Boman HG (2003) Antibacterial peptides: basic facts and emerging concepts. J Intern Med 254: 197–215

    Article  CAS  PubMed  Google Scholar 

  • Diamond G., Zasloff M, Eck H, Brasseur M, Maloy WL, et al. (1991) Tracheal antimicrobial peptide, a cysteine-rich peptide from mammalian tracheal mucosa: peptide isolation and cloning of a cDNA. Proc Natl Acad Sci U S A 88: 3952–3956

    CAS  PubMed  Google Scholar 

  • Frohlich O, Po C, Murphy T, Young LG (2000) Multiple promoter and splicing mRNA variants of the epididymis-specific gene EP2. J Androl 21: 421–430

    CAS  PubMed  Google Scholar 

  • Frohlich O, Po C, Young LG (2001) Organization of the human gene encoding the epididymis-specific EP2 protein variants and its relationship to defensin genes. Biol Reprod 64: 1072–1079

    Article  CAS  PubMed  Google Scholar 

  • Ganz T (2003) Defensins: Antimicrobial peptides of innate immunity. Nat Rev Immunol 3: 710–720

    Article  CAS  PubMed  Google Scholar 

  • Kao CY, Chen Y, Zhao YH, Wu R (2003) ORFeome based search of airway epithelial cell-specific human β-defensin genes. Am J Respir Cell Mol Biol 29: 71–80

    Article  CAS  PubMed  Google Scholar 

  • Kumar S, Tamura K, Jakobsen IB, Nei M (2001) MEGA2: molecular evolutionary genetics analysis software. Bioinformatics 17: 1244–1245

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25: 402–408

    Article  CAS  PubMed  Google Scholar 

  • Lynn DJ, Lloyd AT, O’Farrelly C (2003) In silico identification of components of the Toll-like receptor (TLR) signaling pathway in clustered chicken expressed sequence tags (ESTs). Vet Immunol Immunopathol 93: 177–184

    CAS  PubMed  Google Scholar 

  • Lynn DJ, Higgs R, Gaines S, Tierney J, James T, et al. (2004) Bioinformatic discovery and initial characterisation of nine novel antimicrobial peptide genes in the chicken. Immunogenetics 56: 170–177

    Article  CAS  PubMed  Google Scholar 

  • Nguyen TX, Cole AM, Lehrer RI (2003) Evolution of primate θ-defensins: a serpentine path to a sweet tooth. Peptides 24: 1647–1654

    Article  CAS  PubMed  Google Scholar 

  • Patil A, Hughes AL, Zhang G (2004) Rapid evolution and diversification of mammalian alpha-defensins as revealed by comparative analysis of rodent and primate genes. Physiol Genomics 20: 1–11

    CAS  PubMed  Google Scholar 

  • 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

    CAS  PubMed  Google Scholar 

  • Rodriguez-Jimenez FJ, Krause A, Schulz S, Forssmann WG, Conejo-Garcia JR, et al. (2003) Distribution of new human beta-defensin genes clustered on chromosome 20 in functionally different segments of epididymis. Genomics 81: 175–183

    CAS  PubMed  Google Scholar 

  • Sang Y, Ortega MT, Blecha F, Prakash O, Melgarejo T (2005) Molecular cloning and characterization of three beta-defensins from canine testes. Infect Immun 73: 2611–2620

    CAS  PubMed  Google Scholar 

  • Schutte BC, McCray PB Jr (2002) β-defensins in lung host defense. Annu Rev Physiol 64: 709–748

    Article  CAS  PubMed  Google Scholar 

  • Schutte BC, Mitros JP, Bartlett JA, Walters JD, Jia HP, et al. (2002) Discovery of five conserved β-defensin gene clusters using a computational search strategy. Proc Natl Acad Sci U S A 99: 2129–2133

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Selsted ME, Tang YQ, Morris WL, McGuire PA, Novotny MJ, et al. (1993) Purification, primary structures, and antibacterial activities of beta-defensins, a new family of antimicrobial peptides from bovine neutrophils. J Biol Chem 268: 6641–6648

    CAS  PubMed  Google Scholar 

  • Xiao Y, Hughes AL, Ando J, Matsuda Y, Chen J-F, et al. (2004) A genome-wide screen identifies a single β-defensin gene cluster in the chicken: implications for the origin and evolution of mammalian defensins. BMC Genomics 5: 56

    Article  PubMed  Google Scholar 

  • Yang D, Biragyn A, Hoover DM, Lubkowski J, Oppenheim JJ (2004) Multiple roles of antimicrobial defensins, cathelicidins, and eosinophil-derived neurotoxin in host defense. Annu Rev Immunol 22: 181–215

    Article  PubMed  Google Scholar 

  • Zhang G, Hiraiwa H, Yasue H, Wu H, Ross CR, et al. (1999) Cloning and characterization of the gene for a new epithelial beta-defensin. Genomic structure, chromosomal localization, and evidence for its constitutive expression. J Biol Chem 274: 24031–24037

    CAS  PubMed  Google Scholar 

  • Zhang G, Wu H, Shi J, Ganz T, Ross CR, et al. (1998) Molecular cloning and tissue expression of porcine β-defensin-1. FEBS Lett 424: 37–40

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank Danielle Goodband for her excellent technical support. This work was supported in part by USDA NRICGP grant 2001-35204-10818.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Frank Blecha.

Additional information

The nucleotide sequence data reported in this article have been submitted to GenBank.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sang, Y., Patil, A.A., Zhang, G. et al. Bioinformatic and expression analysis of novel porcine β-defensins. Mamm Genome 17, 332–339 (2006). https://doi.org/10.1007/s00335-005-0158-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00335-005-0158-0

Keywords

Navigation