Characterization of the antimicrobial peptide family defensins in the Tasmanian devil (Sarcophilus harrisii), koala (Phascolarctos cinereus), and tammar wallaby (Macropus eugenii)

Abstract

Defensins comprise a family of cysteine-rich antimicrobial peptides with important roles in innate and adaptive immune defense in vertebrates. We characterized alpha and beta defensin genes in three Australian marsupials: the Tasmanian devil (Sarcophilus harrisii), koala (Phascolarctos cinereus), and tammar wallaby (Macropus eugenii) and identified 48, 34, and 39 defensins, respectively. One hundred and twelve have the classical antimicrobial peptides characteristics required for pathogen membrane targeting, including cationic charge (between 1+ and 15+) and a high proportion of hydrophobic residues (>30%). Phylogenetic analysis shows that gene duplication has driven unique and species-specific expansions of devil, koala, and tammar wallaby beta defensins and devil alpha defensins. Defensin genes are arranged in three genomic clusters in marsupials, whereas further duplications and translocations have occurred in eutherians resulting in four and five gene clusters in mice and humans, respectively. Marsupial defensins are generally under purifying selection, particularly residues essential for defensin structural stability. Certain hydrophobic or positively charged sites, predominantly found in the defensin loop, are positively selected, which may have functional significance in defensin-target interaction and membrane insertion.

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Acknowledgements

This research was supported by the Australian Research Council. We thank Rebecca Johnson, Marc Wilkins, and Peter Timms and other colleagues in the Koala Genome Consortium for providing access to koala sequence data prior to public release.

Authors’ contributions

KB, YC, and DOM conceived and designed the study. EAJ carried out analyses under the supervision of YC and DOM and drafted the paper. KB, YC, and DOM revised the paper.

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Correspondence to Katherine Belov.

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Electronic supplementary material

Supplementary Table 1

Database accession details (PDF 270 kb)

Supplementary Table 2

Tasmanian devil, koala and tammar wallaby defensin and flanking gene scaffold/contig coordinates. Defensin transcription details, previously predicted accession number and protein sequences with predicted mature peptide highlighted in red. (PDF 155 kb)

Supplementary Table 3

Accession numbers and resources for human, mouse, rat, chimpanzee and opossum, sequences used in this study (PDF 194 kb)

Supplementary Table 4

Defensins charge and hydrophobicity characteristics (PDF 89 kb)

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Jones, E.A., Cheng, Y., O’Meally, D. et al. Characterization of the antimicrobial peptide family defensins in the Tasmanian devil (Sarcophilus harrisii), koala (Phascolarctos cinereus), and tammar wallaby (Macropus eugenii) . Immunogenetics 69, 133–143 (2017). https://doi.org/10.1007/s00251-016-0959-1

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Keywords

  • Tasmanian devil
  • Koala
  • Tammar wallaby
  • Defensin
  • Evolution