Molecular Characteristics and Expression Regulation of CABYR in the Testis of Pig

Abstract

CABYR is a protein that regulates calcium-binding tyrosine phosphorylation located in sperm flagella, which increases significantly during sperm capacitation and is primarily involved in sperm capacitation and motility. This study aimed to analyze the expression regulation pattern, sequence characteristics and potential biological function of CABYR gene in Banna mini-pig inbred line (BMI). The testes of adult BMI boars were used for RNA-seq; the complete coding sequence of CABYR was cloned; the sequence, structural characteristics, interacting proteins, KEGG and GO of CABYR were analyzed; the ceRNA regulatory network of CABYR was built using RNA-seq data. The average expression level and TPM value of CABYR gene obtained by RNA-seq was 11 187.50 and 225.29, respectively. The full-length CDS of CABYR was 1149 bp (GenBank accession number: OK042309) encoding 382 amino acids (GenBank login number: UYO37316). The amino acid sequence alignment analysis of multiple species revealed that the similarity between BMI and other species exceeded 73%, indicating a relatively conserved sequence. The CABYR protein was found to contain the DD_CABYR_SP17 conserved domain, the results of species phylogenetic tree analysis met the clustering criteria. Other analyses, including protein-protein interaction (PPI) networks, KEGG, and GO, revealed that BMI CABYR interacts with 50 proteins involved in a variety of functions, such as sexual reproduction and reproductive processes. CABYR was involved in 7 GOs, including five cellular components, one molecular functions, and one biological processes, according to functional annotation. Six miRNAs were found to regulate the CABYR gene via targeted interactions. Additionally, 8, 6, and 1 lncRNAs were found to interact with ssc-miR-4331-3p, ssc-miR-744, and ssc-miR-491, respectively, in association with CABYR. This study examines the expression of CABYR in the testis of BMI, as well as its molecular structure characteristics and expression regulatory network. These findings provide a foundation for future research on the involvement of the CABYR gene in spermatogenesis processes in BMI pigs, particularly in relation to crucial biological processes such as sperm motility and fibrous sheath development.