Evolutionary dynamics and regulatory site analysis of AMP family genes in cattle and sheep

Background: Ruminants possess a rich repository of natural antimicrobial peptides(AMPs) within their bodies, surpassing those found in humans and mice. These peptides, including Defensin, Cathelicidin, and Lysozyme, are integral to the body's innate and adaptive immune responses and represent promising alternatives to antibiotics with significant application potential.

Results: In the present study, we conducted a systematic analysis of 40 Defensins, 38 Cathelicidins, and 61 Lysozymes in cattle and sheep. Our findings revealed that these peptides have retained functional integrity through the evolutionary history of these species. However, they exhibit unique gene duplication and expansion events when compared to humans and mice, indicating their potent roles in cattle and sheep. Notably, the Cathelicidin gene family experienced the most substantial expansion in these ruminants. The newly expanded genes were highly expressed in tissues and organs such as the tongue surface, intestine, mammary gland, and others, exhibiting tissue-specific preferences. This expression pattern is associated with the unique behaviors and high lactation capacity of ruminants. An in vitro bacterial inhibition assay demonstrated that EBD, LALBA, LYSB, and CATHL4 exhibited significant broad-spectrum antibacterial activity. Additionally, loci dB1, dB5, cB2, cB3, and yB1 were pinpointed as key co-regulatory elements in the antimicrobial peptide motifs within cattle mammary epithelial cells.

Conclusions: This research illuminates the structure-function relationship and antimicrobial potency of natural AMP genes in cattle and sheep, providing a theoretical foundation for the development of novel veterinary drugs to treat common bacterial diseases in ruminants and for enhancing animal health care. The identified transcriptional regulatory sites offer a new perspective on the molecular regulation of AMP genes expression, which can be leveraged to improve the disease resistance of domestic animals. This work contributes to a broader understanding of the evolution and regulation of AMP genes, with potential applications for animal health and breeding programs.