Copy number variation of PR-39 cathelicidin, and identification of PR-35, a natural variant of PR-39 with reduced mammalian cytotoxicity

Gene. 2019 Apr 15:692:88-93. doi: 10.1016/j.gene.2018.12.065. Epub 2019 Jan 11.

Abstract

Proline-arginine-rich (PR)-39 is neutrophil antimicrobial peptide that has potent antimicrobial activity against a broad spectrum of microorganisms, including bacteria, fungi, and some enveloped viruses as a part of the innate immune system. We analyzed the nucleotide sequence variations of PR-39 exon 4, which is the mature peptide region responsible for antimicrobial activity, from 48 pigs of six breeds using sequence-based typing. The analysis identified four alleles including allele PR-35 with a 12-bp deletion near the N-terminus. Interestingly, 16.7% of individuals showed the presence of three alleles per individual, but only in the Berkshire and Duroc breeds. We further analyzed the genetic diversity of PR-39 for the entire genomic region of the gene from PR-39 exon 1 to the 3' untranslated region for different alleles by PCR amplification and cloning. The antimicrobial activity of chemically synthesized PR-35 was similar to that of PR-39, but the level of mammalian cell cytotoxicity was lower than the wild type. Better knowledge of the genetic diversity of PR-39 among different individuals and breeds may contribute to improved immune defense of pigs. PR-35, as a natural antimicrobial peptide variant, could be an interesting candidate for the development of peptide antibiotics.

Keywords: Antimicrobial activity; Antimicrobial peptide; Cathelicidin; Cytotoxicity; Sus scrofa.

MeSH terms

  • Animals
  • Antimicrobial Cationic Peptides / adverse effects
  • Antimicrobial Cationic Peptides / genetics
  • Antimicrobial Cationic Peptides / pharmacology*
  • Cathelicidins
  • DNA Copy Number Variations*
  • Drug Evaluation, Preclinical / methods
  • Exons
  • Gene Expression
  • Genome
  • Gram-Negative Bacteria / drug effects
  • HEK293 Cells
  • Humans
  • Microbial Sensitivity Tests
  • Swine
  • Toxicity Tests

Substances

  • Antimicrobial Cationic Peptides
  • PR 39
  • Cathelicidins