Cloning of ACTA1 Gene Promoter Region and Correlation Analysis Between DNA Methylation and Tissue Expression in Yak

doi:10.7668/hbnxb.20190124

Abstract

Abstract: In order to investigate the methylation status and mRNA expression levels of skeletal muscle actin alpha 1(ACTA1)gene in six tissues including muscle and adipose,et al,the experiment was conducted with Leiwuqi yak and Maiwa yak. The bisulfite-sequencing PCR(BSP)was used to detect the methylation pattern of the promoter region of ACTA1 gene,and the ACTA1 gene was detected by Real-time fluorescent quantitative PCR in the gluteus maximus,heart,kidney,lung,liver,and mRNA expression levels in adipose tissue. The results showed that the total length of the partial sequence of the transcription initiation site of the ACTA1 gene and the first exon region of yak was 1 028 bp. The BSP analysis showed that the DNA methylation level of muscle tissue was the lowest,and the methylation rate of adipose tissue was the highest. Among them,the methylation probabilities of Leiwuqi yak gluteus maximus,heart,kidney,lung,liver and adipose were 6.25%,6.88%,20.00%,16.25%,26.25% and 29.38%,methylation probabilities of Maiwa yak gluteus maximus,heart,kidney,lung,liver and adipose were 5.00%,7.50%,18.13%,20.00%,26.25%,28.75%. The expression of ACTA1 gene in kidney, lung, liver and adipose mRNA was significantly lower than that of gluteus maximus (P<0.01), which was significantly lower than that of heart (P<0.05).There was a significant difference in heart mRNA expression between Leiwuqi yak and Maiwa yak (P<0.05), but there was no significant difference in methylation rate and mRNA expression in other tissues of Leiwuqi and Maiwa yak. The methylation rate of lung tissues of Leiwuqi yak is significantly lower than that of Maiwa yak (P<0.05).There was no significant difference in methylation rate and mRNA expression between Leiwuqi and Maiwa yak (P>0.05),and the methylation level of each tissue was significantly negative correlated with the expression of ACTA1 gene mRNA(r=-0.797, P=0.002). Therefore,the results of the study indicate that the DNA methylation pattern of ACTA1 gene has a certain regulatory effect on muscle development,which can provide partial data support for the epigenetic marker of yak genetic breeding.

Key words: Yak, ACTA1 gene, Promoter region, DNA methylation, Bisulfite-sequencing PCR, Tissue expression

CLC Number:

YANG Qin, CHAI Zhixin, WANG Jikun, WANG Hui, ZHONG Jincheng. Cloning of ACTA1 Gene Promoter Region and Correlation Analysis Between DNA Methylation and Tissue Expression in Yak[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(1): 222-229. doi: 10.7668/hbnxb.20190124.

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References

[1] 施奇静,王静,孙军平,薛堃,孟秀祥.中国牦牛资源保护及可持续利用:驯化与品种培育[J].家畜生态学报,2016,37(1):81-86.doi:10.3969/j.issn.1673-1182.2016.01.017.
Shi Q J,Wang J,Sun J P,Xue K,Meng X X. Conservation and sustainable utilization of Yak resources of China:domestication and breed[J].Journal of Domestic Animal Ecology,2016,37(1):81-86.
[2] 朱筱娟,曾宪录,宋朝霞,郝水.细胞核内肌动蛋白及其功能研究进展[J].科学通报,2004,49(11):1031-1035.doi:10.3321/j.issn:0023-074X.2004.11.003.
Zhu Y J,Zeng X L,Song Z X,Hao S. Research progress on actin and its function in nucleus[J]. Chinese Science Bulletin,2004,49(11):1031-1035.
[3] Atencia R,Asumendi A,García-Sanz M. Role of cytoskeleton in apoptosis[J].Vitamins and Hormones,2000,58:267-297.doi:10.1016/S0083-6729(00)58028-5.
[4] Sparrow J C,Nowak K J,Durling H J,Beggs A H,Wallgren-Pettersson C,Romero N,Nonaka I,Laing N G. Muscle disease caused by mutations in the skeletal muscle alpha-actin gene(ACTA1)[J].Neuromuscular Disorders,2003,13(7-8):519-531.doi:10.1016/S0960-8966(03)00101-9.
[5] Gavin R H. Microtubule-microfilament synergy in the cytoskeleton[J].International Review of Cytology,1997,173:207-242.doi:10.1016/S0074-7696(08)62478-X.
[6] Kao J C,Liewluck T,Milone M. A novel ACTA1 mutation causing progressive facios capulo peroneal myopathy in an adult[J].Journal of Clinical Neuroscience,2018,53:261-262.doi:10.1016/j.jocn.2018.04.044.
[7] Ueda K,Serajee F,Huq A M. A mutation in the ACTA1 gene manifesting nemaline myopathy with central nervous system lesions[J]. Journal of Clinical Neurology,2017,13(3):300-302.doi:10.3988/jcn.2017.13.3.300.
[8] Swery C A,Holton J L,Dick D J,Muntoni F,Hanna M G. Zebra body myopathy is caused by a mutation in the skeletal muscle actin gene(ACTA1)[J].Neuromuscul Disord,2015,25(5):388-391.doi:10.1016/j.nmd.2015.02.003.
[9] Sibut V,Hennequet-Antier C,Le Bihan-Duval E,Marthey S,Duclos M J,Berri C. Identification of differentially expressed genes in chickens differing in muscle glycogen content and meat quality[J].BMC Genomics,2011,12(1):112.doi:10.1186/1471-2164-12-112.
[10] Shin S C,Chung E R. Identification of differentially expressed genes between high and low marbling score grades of the Longissimus dorsi muscle in Hanwoo(Korean cattle)[J].Meat Sci,2016,121(5):114-118.doi:10.1016/j.meatsci.2016.05.018.
[11] Guintivano J,Arad M,Gould T D,Payne J L,Kaminsky Z A. Antenatal prediction of postpartum depression with blood DNA methylation biomarkers[J].Molecular Psychiatry,2014,19(5):560-567.doi:10.1038/mp.2013.62.
[12] Dahl C,Guldberg P. DNA methylation analysis techniques[J].Biogerontology,2003,4(4):233-250.doi:10.1023/a:1025103319328.
[13] Vaissière T,Sawan C,Herceg Z. Epigenetic interplay between histone modifications and DNA methylation in gene silencing[J]. Mutat Res,2008,659(1-2):40-48.doi:10.1016/j.mrrev.2008.02.004.
[14] Wilkinson M F. Evidence that DNA methylation engenders dynamic gene regulation[J].Proceedings of the National Academy of Sciences of the United States of America,2015,112(17):E2116.doi:10.1073/pnas.1502195112.
[15] Huang Y Z,Sun J J,Zhang L Z,Li C J,Womack J E,Li Z J,Lan X Y,Lei C Z,Zhang C L,Zhao X,Chen H. Genome-wide DNA methylation profiles and their relationships with mRNA and the microRNA transcriptome in bovine muscle tissue(Bos taurine)[J]. Scientific Reports,2014,4(1):6546.doi:10.1038/srep06546.
[16] 邱峥艳,郭将,张永宏,李传民,赵志辉.莱芜猪与大白猪杂交一代甲基化程度差异分析[J].中国兽医学报,2013,33(5):780-784.doi:10.16303/j.cnki.1005-4545.2013.05.031.
Qiu Z Y,Guo J,Zhang Y H,Li C M,Zhao Z H. Differential analysis of DNA methylation contents in hybirds Yorkshinex Lai wu pig[J].Chinese Journal of Veterinary Science,2013,33(5):780-784.
[17] Wang S,Pan Z,Zhang Q,Xie Z,Liu H,Li Q. Differential mRNA expression and promoter methylation status of SYCP3 gene in testes of Yaks and Cattle-Yaks[J].Reproduction in Domestic Animals,2012,47(3):455-462.doi:10.1111/j.1439-0531.2011.01902.
x.
[18] 刘振山,李齐发,张庆波,屈旭光,董丽艳,强巴央宗,谢庄,刘红林.犏牛及其亲本 IGF2 基因mRNA表达水平、DNA甲基化状态差异分析[J]. 自然科学进展,2009,19(4):380-385.doi:10.3321/j.issn:1002-008X.2009.04.003.
Liu Z S,Li Q F,Zhang Q B,Qu X G,Dong L Y,Qiang B Y Z,Xie Z,Liu H L. Analysis of mRNA expression level and DNA methylation status of IGF2 gene in yak and its parents[J].Advances in Natural Science,2009,19(4):380-385.
[19] 潘增祥,刘振山,李隐侠,于莎莉,李明桂,谢庄,李齐发.犏牛及其亲本睾丸组织中印记基因 SNRPN DMR 甲基化与mRNA表达差异研究[J].中国农业科学,2010,43(22):4709-4716.doi:10.3864/j.issn.0578-1752.2010.22.019.
Pan Z X,Liu Z S,Li Y X,Yu S L,Li M G,Xie Z,Li Q F. Differences of SNRPN methylation status and Its mRNA expression in testes between Cattle-Yaks and their parents[J].Scientia Agricultura Sinica,2010,43(22):4709-4716.
[20] 周阳,骆骅,李伯江,贾超,谢庄,赵兴波,钟金城,李齐发.牦牛和犏牛睾丸组织 DDX4 基因mRNA表达水平与启动子区甲基化[J]. 中国农业科学,2013,46(3):630-638.doi:10.3864/j.issn.0578-1752.2013.03.020.
Zhou Y,Luo H,Li B J,Jia C,Xie Z,Zhao X B,Zhong J C,Li Q F. mRNA expression level and promoter methylation of DDX4 gene in testis of yak and cattle-yak[J].Scientia Agricultura Sinica,2013,46(3):630-638.
[21] 陈洁,李冬杰,刘艳琴,张萃,戴蕴平,李世杰,李宁. 体细胞核移植牛肺脏中 H19 和 Xist 基因的DNA甲基化状态[J].科学通报,2008,53(11):1305-1310.
Chen J,Li D J,Liu Y Q,Zhang C,Dai Y P,Li S J,Li N. DNA methylation status of H19 and Xist genes in bovine lungs after somatic cell nuclear transfer[J].Chinese Science Bulletin,2008,53(11):1305-1310.
[22] Zhou X Y,Liu L L,Jia W C,Pan C Y. Methylation profile of bovine Oct4 gene coding region in relation to three germ layers[J].Journal of Integrative Agriculture,2016,15(3):618-628.doi:10.1016/S2095-3119(15)61100-5.
[23] 赵海谕.奶山羊 PITX1、 PITX2 基因的遗传变异、mRNA表达及DNA甲基化修饰对泌乳性能的影响[D].杨凌:西北农林科技大学,2014.
Zhao H Y. Effects of genetic variation,mRNA expression and DNA methylation of PITX1 and PITX2 genes on lactation performance in dairy goats[D]. Yangling:Northwest A&F University,2014.
[24] 韩玉娇.秦川牛 LCORL 基因的多态性、mRNA表达及启动子区甲基化研究[D]. 杨凌:西北农林科技大学,2016.
Han Y J. Polymorphism,mRNA expression and promoter methylation of LCORL gene in Qinchuan cattle[D]. Yangling:Northwest A&F University,2016.
[25] 房希碧.基于转录组和全基因组甲基化联合分析的牛肉质性状候选基因的鉴定与功能验证[D].长春:吉林大学,2017.
Fang X B. Identification and functional verification of beef trait candidate genes based on transcriptome and genome-wide methylation analysis[D].Changchun:Jilin University,2017.
[26] 姚力丹,祖玲玲,马晓燕,依明·苏来曼,决肯·阿尼瓦什.新疆巴什拜羊肌肉生长抑制素基因DNA甲基化与mRNA表达量分析[J].中国畜牧兽医,2017,45(10):2987-2993.doi:10.16431/j.cnki.1671-7236.2017.10.022.
Yao L D,Zu L L,Ma X Y,Yi M·Su L M,Jue K·A N W. Analysis of DNA methylation and mRNA expression level of MSTN gene in Xinjiang Bashiby sheep[J].China Animal Husbandry & Veterinary Medicine,2017,45(10):2987-2993.
[27] 赵海谕,蓝贤勇,雷初朝,陈宏. 表观遗传学:家畜遗传育种的新挑战[J].家畜生态学报,2014,35(8):1-5.doi:10.3969/j.issn.1673-1182.2014.08.001.
Zhao H Y,Lan X Y,Lei C C,Chen H. Epigenetics:new challenges in animal genetics and breeding[J].Acta Ecologae Animalis Domastici,2014,35(8):1-5.
[28] Ichimura T,Watanabe S,Sakamoto Y,Aoto T,Fujita N,Nakao M. Transcriptional repression and heterochromatin formation by MBD1 and MCAF/AM family proteins[J].Journal of Biological Chemistry,2005,280(14):13928-13935.doi:10.1074/jbc.M413654200.

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