透明带结合蛋白ZPBP2在公猪睾丸中的表达调控及生物信息学分析  

Expression Regulation and Bioinformatics Analysis of Zona Pellucida Binding Protein ZPBP2 in Boars Testis

作　　者：张霞 冯彩平[1] 王瑞[1] 贺莹[1] 王海贞 ZHANG Xia;FENG Caiping;WANG Rui;HE Ying;WANG Haizhen(Department of Biological and Food Engineering,Lyuliang University,Lvliang 033001,Shanxi,China)

机构地区：[1]吕梁学院生物与食品工程系,山西吕梁033001

出　　处：《四川农业大学学报》2025年第2期435-443,共9页Journal of Sichuan Agricultural University

基　　金：吕梁市引进高层次科技人才计划项目(2023RC26);山西省高等学校科技创新项目(2021L571)。

摘　　要：【目的】研究ZPBP2在滇南小耳猪(Diannan small-eared pig,DSE)中的基因结构和蛋白网络调控特征,探寻其在猪受精过程中的生理功能。【方法】采用RNA测序技术对成年DSE的睾丸进行全转录组测序分析,获得ZPBP2在睾丸中的表达量,分析ZPBP2基因结构和蛋白特征;分析氨基酸序列同源性和邻近基因并构建系统进化树;进而挖掘ZPBP2的互作蛋白,并构建蛋白互作网络,进行GO功能注释和KEGG功能富集分析;预测潜在调控的ceRNA并构建调控网络图。【结果】ZPBP2编码区全长981 bp,含有8个外显子以及7个内含子,编码326个氨基酸;二级结构中无规则卷曲的占比最高,含有丝氨酸、苏氨酸和酪氨酸磷酸化位点并且含有信号肽;进化分析表明,ZPBP2在各物种中均具有高度的保守性以及同源性;蛋白互作分析发现ZPBP2与ZP3R、CCT6A、CCT8、ZPBP、CCT3等多种蛋白均存在互作关系;GO与KEGG功能富集分析发现这些蛋白主要富集在受精、精子与透明带的结合、糖解与糖代谢合成以及碳代谢等信号通路上;功能注释发现ZPBP2在免疫球蛋白产生的负向调节、基因表达的昼夜节律调节以及精子与透明带的结合等方面具有重要功能;ceRNA网络图分析表明ZPBP2受ssc-miR-122-5p靶向调控。【结论】ZPBP2基因在DSE受精过程中参与了表达调控,获得的结构特征为进一步研究该基因在猪精子发生过程的功能奠定了基础。【Objective】The study aimed to elucidate the gene structure and protein network characteristics of ZPBP2 in Diannan small-eared pig(DSE)and to explore its physiological function in pig fertilization.【Method】RNA sequencing was employed to conduct a whole-transcriptome analysis of testes from adult DSE pigs.Firstly,the expression level in testes,gene structure,and protein characteristics of ZPBP2 were analyzed.Amino acid sequence homology and neighboring genes were examined,and the phylogenetic tree was constructed.Subsequently,the interacting proteins of ZPBP2 were identified,and the protein interaction network was established.Functional enrichment analyses were performed,including Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG).Finally,regulatory ceRNAs were predicted,and the transcriptional regulatory network was constructed.【Result】The ZPBP2 encoded a protein with a coding region of 981 bp,containing 8 exons and 7 introns,and a polypeptide of 326 amino acids.The secondary structure predominantly featured random coils,including serine,threonine,and tyrosine phosphorylation sites,as well as a signal peptide.Phylogenetic analysis revealed high conservation and homogeneousness of ZPBP2 across species.Protein interaction studies identified that ZPBP2 interacted with ZP3R,CCT6A,CCT8,ZPBP,and CCT3.GO and KEGG analyses found that these proteins were primarily enriched in fertilization,sperm binding to the zona pellucida,glycolysis and carbohydrate metabolism,and carbon metabolism.Functional annotation revealed that ZPBP2 was critical for the negative regulation of immunoglobulin production,the regulation of circadian gene expression,and sperm binding to the zona pellucida.ceRNA network analysis indicated that ZPBP2 was targeted and regulated by ssc-miR-122-5p.【Conclusion】This study revealed that the ZPBP2 gene was involved in the regulation of fertilization in DSE,and its structural characteristics also laid a foundation for further study on the function of ZPBP2 in pig spermatogenesis

关 键 词：猪 透明带结合蛋白2 精子发生 蛋白互作 功能注释 

分 类 号：S828.3[农业科学—畜牧学]