TPPP2基因在保山猪睾丸中的表达、转录调控及潜在的生物学作用机制  

Expression,transcriptional regulation and potential biological mechanism of TPPP2 gene in the testis of Baoshan swine

作　　者：翟兰 席学敏 常勇诚 赵加鼎 富国文 徐世雄 龚绍荣 赵桂英[1] 霍金龙[1] ZHAI Lan;XI Xuemin;CHANG Yongcheng;ZHAO Jiading;FU Guowen;XU Shixiong;GONG Shaorong;ZHAO Guiying;HUO Jinlong(Faculty of Animal Science and Technology,Yunnan Agricultural University,Kunming,Yunnan 650201,China;Baoshan Swine Research Institute,Baoshan,Yunnan 678004,China;College of Animal Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201,China)

机构地区：[1]云南农业大学动物科学技术学院,云南昆明650201 [2]云南省保山市保山猪研究所,云南保山678004 [3]云南农业大学动物医学院,云南昆明650201

出　　处：《福建农林大学学报(自然科学版)》2025年第3期325-334,共10页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：云南省科技厅重大科技专项(202302AE090016);云南省科技厅重点研发项目(2018BB003);云南省施甸县保山猪产业科技特派团项目(202304BI090010);云南省财政厅科教文化专项(A3032022211)。

摘　　要：【目的】研究保山猪睾丸组织TPPP2基因的基本生物学特性及多个组织TPPP2基因的表达水平,为进一步解析TPPP2基因在保山猪中的作用机制提供依据。【方法】通过全转录组测序分析不同月龄保山猪睾丸TPPP2基因的表达量,构建TPPP2 mRNA与miRNA、lncRNA的竞争性内源RNA(ceRNA)调控网络,分析TPPP2的氨基酸同源性及蛋白互作网络,对TPPP2的互作蛋白进行GO、KEGG富集分析,并将获得的互作蛋白与转录组测序所获得的基因表达量进行关联性分析。【结果】TPPP2基因编码序列(CDS)全长516 bp,位于猪第7号染色体上,有4个外显子和3个内含子;蛋白质功能分析发现,TPPP2有171个氨基酸,具有疏水性,含有磷酸化位点;系统进化、同源性和临近基因分析显示,保山猪的TPPP2与多个物种的聚类符合分类学标准,相似度均超过80%,且具有较高保守性的相邻基因。蛋白互作分析显示,TPPP2与OAZ3、NME8蛋白的互作较为密切。KEGG富集分析显示,这些互作蛋白主要富集在T细胞受体信号、鞘脂信号和mRNA监视等通路上。GO富集分析发现,这些蛋白主要富集在精子鞭毛运动、精子DNA凝结、精子能动性、精子核分化等条目上。TPPP2基因受3个miRNA(ssc-miR-150、ssc-miR-205、ssc-miR-222)靶向调控。qPCR检测显示:TPPP2基因在保山猪15个组织中呈现差异表达现象,且在睾丸中的表达量最高;睾丸中TPPP2基因的表达量随着保山猪月龄的增大逐渐上调。【结论】获得了保山猪睾丸组织TPPP2基因的分子特征、蛋白质结构及相互作用转录调控网络,揭示了TPPP2在睾丸中的高表达水平,表明TPPP2基因在猪精子发生中发挥着重要作用。【Objective】This study aimed to investigate the basic biological characteristics of tubulin polymerization promoting protein family member 2(TPPP2)gene in the testis of Baoshan swine,as well as its expression levels in other tissues,so as to provide a basis for elucidating its mechanism in Baoshan swine.【Method】The expression of TPPP2 gene in the testes of Baoshan swine was analyzed at different months through whole-transcriptome sequencing.After constructing a competitive endogenous RNA(ceRNA)regulatory network between TPPP2 mRNA,miRNA,and lncRNA,the amino acid homology and protein interaction network of TPPP2 were analyzed.Moreover,GO and KEGG enrichment analyses were performed on the interacting proteins of TPPP2,and their expression correlation with TPPP2 were analyzed through transcriptome sequencing data.【Result】The TPPP2 gene,located on chromosome 7,had a 516 bp coding sequence(CDS)and consisted of 4 exons and 3 introns.Functional analysis revealed that TPPP2 comprised 171 amino acids,exhibited hydrophobicity,and contained phosphorylation sites.Phylogenetic homology and adjacent gene analyses indicated the clustering of TPPP2 in Baoshan swine with multiple species,which was in accordance with taxonomic standards;with sequence similarity exceeding 80%,it was considered of high conserved neighboring genes.Protein interaction analysis revealed close interaction between TPPP2 and protein OAZ3 and NME8,and these interacting proteins were primarily enriched in pathways involved in T-cell receptor signaling,sphingolipid signaling and mRNA surveillance by KEGG enrichment analysis,and related to sperm flagellum motility,sperm DNA condensation,sperm motility and sperm nuclear differentiation by GO enrichment analysis.Furthermore,TPPP2 was targeted and regulated by 3 miRNA(ssc-miR-150,ssc-miR-205 and ssc-miR-222).qPCR analysis revealed differential expression of TPPP2 across 15 tissues,with the highest expression in testis.Notably,its expression in testicular tissue progressively increased with age in Baos

关 键 词：微管蛋白聚合促进蛋白家族成员2 猪 分子结构 蛋白特征 转录调控 

分 类 号：S828.2[农业科学—畜牧学]