Transcriptional Regulation of Spermatogenesis-Related Gene CDK16 in Banna Mini-Pig Inbred Line
Transcriptional Regulation of Spermatogenesis-Related Gene CDK16 in Banna Mini-Pig Inbred Line
Hongmei Dai1, Shixiong Xu1, Zhipeng Liu1, Hailong Huo2, Fuhua Yang1, Xia Zhang3* and Jinlong Huo1,4*
ABSTRACT
CDK16 is known to have significant involvement in various biological processes of mammalian spermatogenesis. Here, the whole-transcriptome sequencing was performed on the testes of 12-month-old adult boars of Banna mini-pig inbred line (BMI), which identified the CDK16 gene as being highly expressed in the BMI testes. The initial average expression of the CDK16 gene in BMI testes was found to be 4,385, with a corrective average expression value (TPM) of the corresponding transcript ENSSSCT00000035953.3 being 54.3489. The full-length coding region of CDK16 obtained from BMI testes was 1,509 bp (GenBank accession number: OP094608). Gene structure analysis revealed that the CDK16 gene was located on chromosome X of the pig genome and comprised 16 exons. Additionally, Protein structure analysis showed that CDK16 consisted of 502 amino acids with a conserved S_TKc domain. Phylogenetic analysis indicated that the amino acid sequence of CDK16 was highly conserved across mammalian species in evolution. PPI network, KEGG and GO analyses suggested that CDK16 interacted with 50 proteins, with mainly involving in the cell cycle, cellular senescence, p53 signaling pathway, protein kinase activity, phosphorylation, and G1/S transition of mitotic cell cycle. Furthermore, correlation analysis between these proteins and RNA-seq data from BMI testes revealed that CDK16 was significantly associated with CDC6, CDKN1A, BARD1, CCNF, ACTR1B, CDKN1B, CKS2, CCND1, and CCNY. Functional annotations indicated that CDK16 was mainly involved in 9 GO terms, including four molecular functions (MF), three cellular components (CC), and two biological processes (BP). Furthermore, the ceRNA network analysis showed that BMI CDK16 was primarily regulated by six miRNAs including ssc-miR-21-3p, ssc-miR-296-3p, ssc-miR-296-5p, ssc-miR-370, ssc-miR-490 and ssc-miR-532-3p. Overall, our findings revealed the crucial role of CDK16 in BMI spermatogenesis, representing a promising candidate for further investigation.
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