Abstract：Abstract The ATP synthase subunit beta (ATP5B) is one of key factors in ATP synthesis, participating in various metabolic activities. Nanyang cattle (Bos taurus) is the one of the five major Chinese native cattle varieties, famous for its adaptability. This study investigated the genetic variation in ATP5B promoter regions and the association of Nanyang cattle growth traits; estimated the conservation of ATP5B among different species, and examined the mRNA expression. It was sequenced the ATP5B promoter region to determine the genetic variants and their association with Nanyang cattle growth traits. To find the reason of this association, It was detected the promoter activity of each genotype promoter at these SNPs site using dual luciferase assay. In the results, there were 14 similar motifs and 5 conservative functional domains in the ATP5B of 7 species, such as, Bos taurus, Sus scrofa, Ruttus and so on. The ATP5B was widely expressed in many tissues in Nanyang cattle, and the muscle tissue had the significant high mRNA level than other tissues (P＜0.05), the ATP5B mRNA in fat tissue was following, the next was in heart, kidney, lung, and liver; the ATP5B mRNA in spleen was the lowest compared with other tisuses (P＜0.05). 3 SNPs in ATP5B promoter regions were identified, named as g.-428T>A, g.-390T>C and g.-322C>G, respectively. After association analysis, there were 3 mutations fell into Hardy-Weinberg equilibrium significantly (Chi-Squared test, P＞0.05). The polymorphism information content (PIC) classification indicated that the 3 SNPs were moderately polymorphic (0.25＜PIC＜0.50). At g.-428T>A locus, individuals with TT genotype exhibited significantly higher body weight, body height and chest circumference than those with AA genotype (P＜0.01). Such as, TA individuals had greater body weight than AA individuals (P＜0.01), but not than TT individuals (P＞0.05). In body height, the individuals with TT or TA had greater level than AA individuals (P＜0.05). In chest circumference, the TT individuals had greater level than the individuals with TA or AA (P＜0.01). So, the TT was the preferable genotype. At g.-390T>C locus, the body height and chest circumference of animals with TT genotype were significantly higher (P＜0.01) than these traits of animals with CC genotype. TC individuals had greater body height than CC individuals (P＜0.05), but less than TT individuals (P＜0.01). TC individuals had less chest circumference than TT individuals (P＜0.01). So, the TT was the preferable genotype. Moreover, the g.-322C>G locus were significantly affected body weight and body length (P＜0.01 or P＜0.05), including that, the CC individuals had greater body weight than CG individuals (P＜0.05) and GG individuals (P＜0.01), meanwhile, CG individuals had greater body weight than GG individuals (P＜0.05). In body length, the individuals with CC or CG had greater level than GG individuals (P＜0.05). So that, CC was the preferable genotype. Last, according to the report of dual luciferase assay, although there were many difference between each promoter style at each SNP site, but all of them not reached at significant level (P＞0.05). The molecular mechanism of regulation between these SNPs and growth traits need to further search. To sum up, these results suggested that the SNPs of ATP5B promoter region, the TT at -428T>A locus, TT at -390T>C locus, and CC at -322C>G locus, affected the growth traits of Nanyang cattle, and they could be used as candidate genotype for breeding programs.
[1]胡建新, 何坚, 刘军, 等.氧自由基对生殖细胞三磷酸腺苷酶和生长抑素的影响[J].中国组织工程研究与临床康复, 2011, (18):3361-3364
[2]肖凯, 王庆松.PTSD样情感行为异常大鼠脑组织ATP酶活性改变[J].西南国防医药, 2002, 12(3):204-207
[3]Chaze T, Meunier B, Chambon C, et al.In vivo proteome dynamics during early bovine myogenesis[J].PROTEOMICS, 2008, 8(20):4236-4248
[4]Neckelmann N, Warner C K, Chung A, et al.The human ATP synthase beta subunit gene: sequence analysis, chromosome assignment, and differential expression[J].GENOMICS, 1989, 5(4):829-843
[5]Xu H, Xu Y, Liang X, et al.Porcine skeletal muscle differentially expressed gene ATP5B : molecular characterization, expression patterns, and association analysis with meat quality traits[J].MAMMALIAN GENOME, 2013, 24(3-4):142-150
[6]段美艳, 赵志东, 李安宁, 等.牛ATP5B基因启动子与组织差异性表达研究[J].畜牧兽医学报, 2015, 46(3):382-387
[7]段美艳, 李安宁, 赵志东, 等.牛ATP5B基因启动子双荧光素酶报告基因载体构建与活性检测[J].西北农林科技大学学报自然科学版, 2015, 43(8):39-45
[8]Garieri M, Delaneau O, Santoni F, et al.The effect of genetic variation on promoter usage and enhancer activity[J].Nature Communications, 2017, 8(1):9-
[9]Reggiani C, Coppens S, Sekhara T, et al.Novel promoters and coding first exons in DLG2 linked to developmental disorders and intellectual disability[J].Genome Medicine, 2017, 9(67):20-
[10]Zhang H, Lu Y, Sun G, et al.The common promoter polymorphism rs11666254 downregulates FPR2/ALX expression and increases risk of sepsis in patients with severe trauma[J].CRITICAL CARE, 2017, 21(171):12-
[11]Pisignano G, Napoli S, Magistri M, et al.A promoter-proximal transcript targeted by genetic polymorphism controls E-cadherin silencing in human cancers[J].Nature Communications, 2017, 8:16-
[12]Yuan X, Cheng G, Yu J, et al.The TERT promoter mutation incidence is modified by germline TERT rs2736098 and rs2736100 polymorphisms in hepatocellular carcinoma[J].Oncotarget, 2017, 8(14):23120-23129
[13]Li Y, Bao C, Gu S, et al.Associations between novel genetic variants in the promoter region of MALAT1 and risk of colorectal cancer[J].Oncotarget, 2017, 8(54):92604-92614
[14]Saitou N, Nei M.The neighbor-joining method: a new method for reconstructing phylogenetic trees[J].Molecular Biology & Evolution, 1987, 4(4):406-
[15]Felsenstein J.Confidence Limits on Phylogenies: An Approach Using the Bootstrap[J].Evolution, 1985, 39(4):783-791
[16]Zuckerkandl E, Pauling L.Evolutionary Divergence and Convergence in Proteins[J].Evolving Genes & Proteins, 1965, 97:97-166
[17]Kumar S, Stecher G, Tamura K.MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets[J].Molecular Biology & Evolution, 2016, 33(7):1870-
[18]Bailey T L, Boden M, Buske F A, et al.MEME Suite: tools for motif discovery and searching[J].NUCLEIC ACIDS RESEARCH, 2009, 37(Web Server issue):W202-W208
[19]Marchler-Bauer A, Bo Y, Han L, et al.CDD/SPARCLE: functional classification of proteins via subfamily domain architectures[J].NUCLEIC ACIDS RESEARCH, 2017, 45(D1):D200-
[20]Marchlerbauer A, Derbyshire M K, Gonzales N R, et al.CDD: NCBI's conserved domain database[J].NUCLEIC ACIDS RESEARCH, 2015, 43(Database issue):D222-
[21]Marchler-Bauer A, Lu S, Anderson J B, et al.CDD: a Conserved Domain Database for the functional annotation of proteins[J].NUCLEIC ACIDS RESEARCH, 2011, 39(Database issue):D225-
[22]田万强, 梅楚刚, 付常振, 等.牛PRKAA2基因SNP检测及与其生长和肉质性状的关联分析[J].西北农林科技大学学报:自然科学版, 2016, 44(7):1-9
[23]李圣彦, 郎志宏, 黄大昉.真核生物启动子研究概述[J].生物技术进展, 2014, (2):158-164
[24]赵卫华, 孟凡义, 赖永榕, 等.急性白血病死亡相关蛋白激酶基因启动子的甲基化[J].南方医科大学学报, 2017, (3):407-410
[25]Alsiddig M A, Yu S G, Pan Z X, et al.Association of single nucleotide polymorphism in melatonin receptor 1A gene with egg production traits in Yangzhou geese[J].ANIMAL GENETICS, 2017, 48:-
[26]Huang Y Z, Li M X, Wang J, et al.A 5′- Regulatory Region and Two Coding Region Polymorphisms Modulate Promoter Activity and Gene Expression of the Growth Suppressor Gene ZBED6 in Cattle[J].PLoS One, 2013, 8(11):e79744-
[27]Cosenza G, Iannaccone M, Pico B A, et al.Molecular characterisation, genetic variability and detection of a functional polymorphism influencing the promoter activity of OXT gene in goat and sheep[J].JOURNAL OF DAIRY RESEARCH, 2017, 84(2):165-
[28]Liu Z, Sun W, Zhao Y, et al.The effect of variants in the promoter of BMPER on the intramuscular fat deposition in longissimus dorsi muscle of pigs[J].GENE, 2014, 542(2):168-172
[29]戴亚丽, 蔡德鸿, 张桦, 等.抑癌基因RAS启动子甲基化与乳头状甲状腺癌的关系[J].首都医科大学学报, 2011, 32(1):135-138
