绵羊Myostatin基因5′调控区的生物信息学分析及孕激素对调控区活性的影响

doi:10.3864/j.issn.0578-1752.2012.09.017

Abstract

Abstract: 【Objective】 The aim of this study is to clone and analyze the 5′ regulatory region of myostatin gene in sheep and to investigate the effect of progesterone on the activity of the 5′ regulatory region. 【Method】 The 5′ regulatory region of myostatin gene in sheep was cloned by PCR, and the regulatory motifs and transcriptional factors, along the region or the similar regions of bovine and porcine, were analyzed and compared by some softwares such as MatInspector. The EGFP was selected as reporter gene and the expression vectors were constructed. The vector was transfected into C2C12 cells and the different doses of progesterone (0 nmol•L-1, 10 nmol•L-1, 100 nmol•L-1 and 1 000 nmol•L-1) were supplemented, then the mRNA of reporter gene EGFP or endogenous myostatin gene was detected by RT-PCR. 【Result】The 5′ regulatory region of myostatin gene in sheep was obtained, and two sites that is possibly contributive to double-muscular character were predicted. Many putative regulatory motifs and transcriptional factors, which may play important roles in the transcriptional regulation of myostatin gene, were found along the regulatory region of sheep, bovine and porcine, such as E-box, MEF2, MTATA, MTBF, HOMF, PRE, ARE and GRE. Most of motifs were found in three animals, but the bases, positions and numbers of the motifs were different among the three animals. The expression vectors pMSTN5’Regu-EGFP of sheep were successfully constructed and transfected. 100 nM progesterone obviously inhibited the mRNA of reporter gene EGFP and endogenous myostatin gene. 【Conclusion】The transcription of myostatin gene in sheep was regulated by various transcriptional factors and corresponding regulatory motifs, and the appropriate dose of progesterone inhibited the transcription of myostatin gene by downregulating the activity of the myostatin gene 5′ regulatory region.

Key words: sheep, Myostatin gene 5&prime, regulatory region, bioinformatic analysis, regulatory motif, transcriptional factor, progesterone

QIN Jian, DU Juan, YANG Ya-Qun, DU Rong. Bioinformatic Analysis of Myostatin Gene 5′ Regulatory Region from Sheep and Effect of Progesterone on the Activity of the Regulatory Region[J].Scientia Agricultura Sinica, 2012, 45(9): 1814-1825.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2012.09.017

https://www.chinaagrisci.com/EN/Y2012/V45/I9/1814

References

[1]McPherron A C, Lawler A M, Lee S J. Regulation of skeletal muscle mass in mice by a new TGF-β superfamily member. Nature, 1997, 387: 83-90.

[2]Zimmers T A, Davies M V, Koniaris L G, Haynes P, Esquela A F, Tomkinson K N, McPherron A C, Wolfman N M, Lee S J. Induction of cachexia in mice by systemically administered myostatin. Science, 2002, 296(5572): 1486-1488.

[3]McPherron A C, Lee S J. Double muscling in cattle due to mutations in the myostatin gene. Proceedings of the National Academy of Sciences of the USA, 1997, 94(23): 12457-12461.

[4]Spiller M P, Kambadur R, Jeanplong F, Thomas M, Martyn J K, Bass J J, Sharma M. The myostatin gene is a downstream target gene of basic helix-loop-helix transcription factor MyoD. Molecular and Cellular Biology, 2002, 22(20): 7066-7082.

[5]杜荣, 安晓荣, 陈永福, 秦健. 绵羊Myostatin 基因启动子的功能分析. 中国科学C辑: 生命科学, 2007, 37(5): 551-557.

Du R, An X R, Chen Y F, Qin J. Functional analysis of the myostatin gene promoter in sheep. Science in China Series C: Life Sciences, 2007, 37(5): 551-557. (in Chinese)

[6]Ma K, Mallidis C, Artaza J, Taylor W, Gonzalez-cadavid N, Bhasin S. Characterization of 5′ regulatory region of human myostatin gene: regulation by dexamethasone in vitro. American Journal of Physiology-Endocrinology and Metabolism, 2001, 281(6): E1128-E1136.

[7]Salerno M S, Thomas M, Forbes D, Watson T, Kambadur R, Sharma M. Molecular analysis of fiber type-specific expression of murine myostatin promoter. The American Journal of Physiology-Cell Physiology, 2004, 287(4): 1031-1040.

[8]Allen D L, Unterman T G. Regulation of myostatin expression and myoblast differentiation by FoxO and SMAD transcription factors. The American Journal of Physiology - Cell Physiology, 2007, 292(1): C188-C199.

[9]于灵芝, 唐辉, 王继英, 武英, 邹丽丽, 姜运良, 吴常信, 李宁. 肌肉生长抑制素基因5′调控区的多态性与大白猪的早期生长有关. 中国科学C辑: 生命科学, 2007, 37(4): 435-440.

Yu L Z, Tang H, Wang J Y, Wu Y, Zou L L, Jiang Y L, Wu C X, Li N. Polymorphisms in the 5′ regulatory region of myostatin gene are associated with early growth traits in Yorkshire pigs. Science in China Series C: Life Sciences, 2007, 37(4): 435-440. (in Chinese)

[10]于政权, 李岩, 孟庆勇, 袁晶, 赵志辉, 李慰, 颜炳学, 于舒洋, 胡晓湘, 樊宝良, 李宁. 利用比较基因组学方法对猪BAC DNA序列分析来研究抑肌素基因的调控. 中国科学C辑: 生命科学, 2004, 34(4): 364-375.

Yu Z Q, LI Y, Meng Q Y, Yuan J, Zhao Z H, Li W, Yan B X, Yu S Y, Hu X X, Fan B L, Li N. Comparative analysis of the pig BAC sequence involved in the regulation of myostatin gene. Science in China Series C: Life Sciences, 2004, 34(4): 364-375. (in Chinese)

[11]顾志良, 卢祥云, 朱大海, 李辉. 鹅肌肉生长抑制素基因5′调控区序列特征和组织表达分析. 畜牧兽医学报, 2008, 39(11): 1606-1611.

Gu Z L, Lu X Y, Zhu D H, Li H. Characterization of 5′ regulatory region and expressional distribution of myostatin gene in goose. Acta Veterinaria et Zootechnica Sinica, 2008, 39(11): 1606-1611. (in Chinese)

[12]Du R, Chen Y F, An X R, Yang X Y, Ma Y, Zhang L, Yuan X L, Chen L M, Qin J. Cloning and sequence analysis of myostatin promoter in sheep. DNA Sequence, 2005, 16(6): 412-417.

[13]Du R, An X R, Chen Y F, Qin J. Some motifs were important for myostatin transcriptional regulation in sheep. Journal of Biochemistry and Molecular Biology, 2007, 40(4): 547-553.

[14]Du R, Du J, Qin J, Cui L C, Hou J, Guan H, An X R. Molecular cloning and sequence analysis of the cat myostatin 5′ regulatory region. African Journal of Biotechnology, 2011, 10(51): 10366-10372.

[15]Apone S, Hauschka S D. Muscle gene E-box control elements. Evidence for quantitatively different transcriptional activities and the binding of distinct regulatory factors. The Journal of Biological Chemistry, 1995, 270: 21420-21427.

[16]Ceccarelli E, McGrew M J, Nguyen T, Grieshammer U, Horgan D, Hughes S H , Rosenthal N. An E box comprises a positional sensor for regional differences in skeletal muscle gene expression and methylation. Developmental Biology, 1999, 213(1): 217-229.

[17]Olson E N, Perry M, Schulz R A. Regulation of muscle differentiation by the MEF2 family of MADS box transcription factors. Developmental Biology, 1995, 172(1): 2-14.

[18]Akkila W M, Chambers R L, Ornatsky O I, McDermott J C. Molecular cloning of up-regulated cytoskeletal genes from regenerating skeletal muscle: potential role of myocyte enhancer factor 2 proteins in the activation of muscle-regeneration-associated genes. Biochemical Journal, 1997, 325: 87-93.

[19]Clop A, Marcq F, Takeda H, Pirottin D, Tordoir X, BibéB, Bouix J, Caiment F, Elsen J M, Eychenne F, Larzul C, Laville E, Meish F, Milenkovic D, Tobin J, Charlier C, Georges M. A mutation creating a potential illegitimate microRNA target site in the myostatin gene affects muscularity in sheep. Nature Genetics, 2006, 38: 813-818.

[20]Ciarmela P, Wiater E, Smith S M, Vale W. Presence, actions, and regulation of myostatin in rat uterus and myometrial cells. Endocrinology, 2009, 150(2): 906-914.

[21]Forde N, Carter F, Fair T, Crowe M A, Evans A C O, Spencer T E, Bazer F W, McBride R, Boland M P, O’Gaora P, Lonergan P, Roche J F. Progesterone-regulated changes in endometrial gene expression contribute to advanced conceptus development in cattle. Biology of Reproduction, 2009, 81(4): 784-794.

[22]Wong C L, Huang Y Y, Ho W K, Poon H K, Cheung P L, O Sum W, Chow P H. Growth-differentiation factor-8 (GDF-8) in the uterus: its identification and functional significance in the golden hamster. Reproductive Biology and Endocrinology, 2009, 7: 134-146.

[23]Ciarmela P, Bloise E, Gray P C, Carrarelli P, Islam S, de Pascalis F, Severi F M, Vale W, Castellucci M, Petraglia F. Activin-A and myostatin response and steroid regulation in human myometrium: disruption of their signaling in uterine fibroid. The Journal of Clinical Endocrinology and Metabolism, 2011, 96(3): 755-765.

Related Articles 15

[1]	LIU YuFang,CHEN YuLin,ZHOU ZuYang,CHU MingXing. miR-221-3p Regulates Ovarian Granulosa Cells Apoptosis by Targeting BCL2L11 in Small-Tail Han Sheep [J]. Scientia Agricultura Sinica, 2022, 55(9): 1868-1876.
[2]	CHE DaLu,ZHAO LiChen,CHENG SuCai,LIU AiYu,LI XiaoYu,ZHAO ShouPei,WANG JianCheng,WANG Yuan,GAO YuHong,SUN XinSheng. Effect of Litter Bed on Growth Performance and Odor Emission in Fattening Lamb [J]. Scientia Agricultura Sinica, 2022, 55(24): 4943-4956.
[3]	SONG ShuZhen, GAO LiangShuang, LI Hong, GONG XuYin, LIU LiShan, WEI YuBing. Effects of Feeding Levels on Muscle Tissue Structure and Muscle Fiber Composition Related Genes in Sheep [J]. Scientia Agricultura Sinica, 2022, 55(21): 4304-4314.
[4]	ChunTao ZHANG,Tao MA,Yan TU,QiYu DIAO. Effects of Circadian Rhythm on Rumen Fermentation and Nutrient Digestion of Mutton Sheep [J]. Scientia Agricultura Sinica, 2022, 55(18): 3664-3674.
[5]	LIU WangJing,TANG DeFu,AO ChangJin. Effect of Allium mongolicum Regel and Its Extracts on the Growth Performance, Carcass Characteristics, Meat Quality and Serum Biochemical Indices of Captive Small-Tailed Han Sheep [J]. Scientia Agricultura Sinica, 2022, 55(17): 3461-3472.
[6]	LIANG Peng,ZHANG TianWen,MENG Ke,SHAO ShunCheng,ZOU ShiFan,RONG Xuan,QIANG Hao,FENG DengZhen. Association Analysis of the ADIPOQ Variation with Sheep Growth Traits [J]. Scientia Agricultura Sinica, 2022, 55(11): 2239-2256.
[7]	KE Na,HAO ZhiYun,WANG JianQing,ZHEN HuiMin,LUO YuZhu,HU Jiang,LIU Xiu,LI ShaoBin,ZHAO ZhiDong,HUANG ZhaoChun,LIANG WeiWei,WANG JiQing. The miR-221 Inhibits the Viability and Proliferation of Ovine Mammary Epithelial Cells by Targeting IRS1 [J]. Scientia Agricultura Sinica, 2022, 55(10): 2047-2056.
[8]	WANG Qian,LI Zheng,ZHAO ShanShan,QIE MengJie,ZHANG JiuKai,WANG MingLin,GUO Jun,ZHAO Yan. Application of Stable Isotope Technology in the Origin Traceability of Sheep [J]. Scientia Agricultura Sinica, 2021, 54(2): 392-399.
[9]	LI SongMei,QIU YuGe,CHEN ShengNan,WANG XiaoMeng,WANG ChunSheng. CRISPR/Cas9 Mediated Exogenous Gene Knock-in at ROSA26 Locus in Sheep Umbilical Cord Mesenchymal Stem Cells [J]. Scientia Agricultura Sinica, 2021, 54(2): 400-411.
[10]	WANG Chen,ZHANG HongWei,WANG HuCheng,SUN XiaoPing,LI FaDi,YANG BoHui. Energy and Protein Requirements of Alpine Merino Growing Sheep [J]. Scientia Agricultura Sinica, 2021, 54(16): 3537-3548.
[11]	WANG JiQing,HAO ZhiYun,SHEN JiYuan,KE Na,HUANG ZhaoChun,LIANG WeiWei,LUO YuZhu,HU Jiang,LIU Xiu,LI ShaoBin. Screening, Identification and Functional Analysis of Important LncRNAs for Lactation Traits in Small-Tailed Han Sheep [J]. Scientia Agricultura Sinica, 2021, 54(14): 3113-3123.
[12]	ZHANG Wei,WANG ShiYin,GAO Li,YANG LiWei,DENG ShuangYi,LIU XiaoNa,SHI GuoQing,GAN ShangQuan. Investigation of miR-486 Target Genes in Skeletal Muscle of Bashbay Sheep in Different Development Periods [J]. Scientia Agricultura Sinica, 2021, 54(14): 3134-3148.
[13]	LI RunTing,CHEN LongXin,ZHANG LiMeng,HE HaiYing,WANG Yong,YANG RuoChen,DUAN ChunHui,LIU YueQin,WANG YuQin,ZHANG YingJie. Transient Expression and the Effect on Proliferation and Apoptosis of Granule Cell Stimulating Factor in Ovarian Fibroblasts [J]. Scientia Agricultura Sinica, 2021, 54(11): 2434-2444.
[14]	LI WenJuan,TAO Hui,ZHANG NaiFeng,MA Tao,DIAO QiYu. Effects of High-Fat Diet on Energy Metabolism and Slaughter Performance of Early-Weaning Lambs [J]. Scientia Agricultura Sinica, 2021, 54(10): 2206-2216.
[15]	SHI TianPei,WANG XinYue,HOU HaoBin,ZHAO ZhiDa,SHANG MingYu,ZHANG Li. Analysis and Identification of circRNAs of Skeletal Muscle at Different Stages of Sheep Embryos Based on Whole Transcriptome Sequencing [J]. Scientia Agricultura Sinica, 2020, 53(3): 642-657.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

Bioinformatic Analysis of Myostatin Gene 5′ Regulatory Region from Sheep and Effect of Progesterone on the Activity of the Regulatory Region

PDF

Abstract

Cite this article

share this article

References

Related Articles 15

Metrics

Comments

Recommended 0