Purpose To study the sequence features, molecular structure, and expression regulation patterns of glutathione peroxidase 4 (GPX4) gene in Baoshan pigs (BS).
Methods The expression level of GPX4 was quantized based on transcriptome sequencing results of BS boar’s testis, and the primary, secondary, and tertiary structure features of GPX4 protein were analyzed. The multi-species amino acid sequence evolutionary tree and protein interaction network were constructed to analyze the conservative homology of GPX4 among various species and the functions of interacting proteins. A competing endogenous RNA (ceRNA) regulatory network diagram of GPX4 was constructed, and KEGG and GO annotations were performed.
Results The coding sequence of GPX4 gene was 594 bp, encoding 197 amino acids, located on No.2 chromosome of pig, including seven exons and six introns. The hydrophobicity analysis results showed that both the N-terminal and C-terminal regions of the protein were hydrophobic. The largest proportion of random coil was observed in the secondary and tertiary structures, accounting for 36.65%. The systematic evolutionary analysis indicated that the amino acid sequence of GPX4 was relatively conserved among mammals, particularly showed a closer phylogenetic relationship with goats and sheep within the Artiodactyla order. The protein interaction network identified several proteins that interacted with GPX4, such as GSR, PRDX6, GRSF1, CHAC1, and HSPA5. These proteins were mainly involved in the pathways related to thyroid hormone synthesis, platinum drug resistance, hepatocellular carcinoma, ferroptosis, and so on. Gene function analysis found that GPX4 gene mainly participated in the physiological processes such as spermatogenesis, antioxidation, and resistance to ferroptosis. The ceRNA regulatory network analysis revealed that the BS GPX4 gene was targeted and regulated by the ssc-miR-214-3p. The qPCR results showed the relative expression level of GPX4 was the highest in the testis of BS, and its expression increased with the age of BS.
Conclusion The results lay the groundwork for a deeper investigation into the transcriptional regulatory role of GPX4 in the spermatogenesis process of BS.
DownLoad: