Abstract
BackgroundThe function of cathelicidins-4 (CATH4) is not limited to microbial killing, but extends to other aspects of immunity and tissue repair. The presence of different CATH4 variants including intron retention affects the immunity system. Intron retention, in buffalo, is not fully studied. In this study, we investigated CATH4 mRNA in river buffalo and their variants, which can be used in the future for selecting buffalo resistant to diseases.Results and conclusionAnalysis of CATH4 mRNA in river buffalo (Egyptian breed) revealed the presence of a novel variant (1073 bp) which includes unspliced part of intron 3 (469 bp) in addition to previously reported unspliced complete intron 1 (103) and intron 2 (137 bp). Identification of intron retention was conducted by comparing the amplified unspliced cDNA and DNA sequences. Analysis of the 3 retained intronic regions revealed the presence of the 4 splice signals, needed for splicing which include the 5′ (GT) and 3′ (AG) intron splice sites, the branch point, and the polypyrimidine tract. However, in the intron-retained sequence, the polypyrimidine tract was weak. It contained 6 and 4 non-continuous uridine stretch in introns 1 and 2, respectively, (intron 3 was partial) which may have caused introns retention. In addition, analysis of the unspliced sequence showed three unique exonic SNPS located close to the splice sites (1 to 22 nucleotides) and five SNPs in retained intronic regions located near the splice sites (18 to 246 nucleotides away from exon/intron boundaries) which may be related to the retention of the three introns.
Highlights
The function of cathelicidins-4 (CATH4) is not limited to microbial killing, but extends to other aspects of immunity and tissue repair
We investigated the different splice variants of mRNA CATH4 in river buffalo (Egyptian breed) which can be used for selecting disease-resistant breeds of buffalo
In three samples that the first primer pairs resulted in 526 bp amplicon which corresponded to CATH4 full-spliced coding region with complete 4 exons and part of 3′ untranslated region (Fig. 1)
Summary
The function of cathelicidins-4 (CATH4) is not limited to microbial killing, but extends to other aspects of immunity and tissue repair. The presence of different CATH4 variants including intron retention affects the immunity system. We investigated CATH4 mRNA in river buffalo and their variants, which can be used in the future for selecting buffalo resistant to diseases. The function of cathelicidins is not limited to microbial killing, but extends to other aspects of immunity and tissue repair (Gallo et al 2002). CATH4 contains 4 exons and 3 introns: the first 3 exons comprise signal peptide and cathelin prodomain (N-terminal) while the fourth exon encodes the cleavage site and variable C-terminal antimicrobial peptide (Zanetti et al 2000; Zaiou and Gallo 2002). Buffalo surpass the cattle in its ability to adapt to the hot, humid areas of muddy and swampy lands
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