基于转录组测序分析SPACA1在版纳微型猪近交系睾丸中的表达及转录调控特征

Transcriptomic analysis of expression and transcriptional regulation of SPACA1 in the testes of Banna mini-pig inbred line

旨在分析版纳微型猪近交系(Banna mini-pig inbred line,BMI)SPACA1基因的序列特征、生物学功能及在睾丸中的表达调控模式。选取4头成年BMI公猪的睾丸进行全转录组测序,获得SPACA1基因在睾丸中的表达丰度。采用RT-PCR技术克隆SPACA1的完整编码序列,分析SPACA1的序列、染色体定位、分子特征及相应蛋白的结构和功能。对SPACA1的相互作用蛋白进行GO、KEGG功能富集,并将获得的蛋白与转录组测序所获得的基因表达量进行关联分析。利用转录组测序数据构建SPACA1 mRNA与miRNA和lncRNA的ceRNA调控网络。SPACA1基因CDS区全长为888 bp(Genbank登录号:OK042308),具有2个跨膜螺旋结构,与SPACA3、SPACA4、HIPK4和CCDC62显著相关;SPACA1及其互作蛋白主要参与精子细胞发育、分化,精子活力、顶体反应和精子鞭毛形成等通路;SPACA1基因受6个miRNAs靶向调控。本研究报道了SPACA1的表达、分子结构、蛋白质功能、在睾丸中的和转录调控作用,为深入研究SPACA1基因在BMI受精以及顶体反应等重要生物学过程中的功能奠定基础。

This study analyzed the sequence characteristics, biological functions and expression regulation pattern of SPACA1 gene in the testis of Banna mini-pig inbred line(BMI). The testis samples of four adult BMI boars were collected for whole transcriptome sequencing to obtain the expression abundance of SPACA1 gene. The complete coding sequence of SPACA1 was cloned by RT-PCR, and the sequence, chromosome location, molecular characteristics, structural and functions of the corresponding protein were analyzed. The interacting proteins of SPACA1 were enriched using GO and KEGG, and the correlation analysis was performed between the proteins and the gene expression abundance. Transcriptome data were used to construct the ceRNA regulatory network among m RNA, miRNA and lncRNA of SPACA1. The full-length CDS of SPACA1 was 888 bp(Genbank accession number:OK042308), and SPACA1 protein contained two transmembrane helix structures. SPACA1 significantly correlated with SPACA3, SPACA4, HIPK4 and CCDC62. SPACA1 and its interacting proteins were mainly involved in germ cell development, spermatid differentiation, sperm motility, acrosomal vesicle and sperm flagellum. BMI SPACA1gene was targeted by six miRNAs. Overall, our study provided comprehensive information on the expression,molecular structure characteristics, protein functions, and transcriptional regulation of SPACA1 in testis, laying a foundation for further study on the functions of SPACA1 gene in important biological processes such as BMI fertilization and acrosome reaction.