牛MSTN启动子与MEF2C转录因子相互作用的研究

doi:10.11843/j.issn.0366-6964.2019.08.005

Abstract

Abstract: The aim of this study was to explore the regulation mechanism of MSTN on muscle growth and development by analysis of the interaction regulation between the MSTN promoter and MEF2C in cattle. Firstly, the 2 267 bp upstream promoter region of MSTN and 1 425 bp coding region of MEF2C were amplified by PCR. Secondly, pGL3-Basic-MSTN and pcDNA3.1(+)-MEF2C dual luciferase reporter vectors were constructed and co-transfected into C2C12 myoblasts by liposome transfection method. The dual luciferase activity were measured after 24 hours. Finally, the core fragment with high activity of MSTN promoter was amplified by PCR. The recombinant expression vector pEGFP-N3-MSTN-P1-MEF2C which replaced (CMV) area with MSTN-P1 fragment was reconstructed, and it was transiently transfected into C2C12 myoblasts and rat mandibular gland epithelial cells, respectively, the expression of green fluorescent protein in cells was observed after 24 hours. Total RNA was extracted from cells after 48 hours. The expression levels of MEF2C in different cells were detected by qRT-PCR.The results showed that the activity of MSTN promoter was enhanced significantly by co-transfected of pcDNA3.1(+)-MEF2C(P<0.05), and it was enhanced extremely significantly compared with pGL3-Basic in C2C12 myoblasts(P<0.01); MSTN-P1 promoter core fragment could drive the expression of MEF2C, and the mRNA expression levels were extremely significantly up-regulated(P<0.01) in C2C12 myoblasts, and significantly up-regulated(P<0.05) in rat mandibular gland epithelial cells. The results indicated that both MSTN promoter and MEF2C could be involved in the regulation of growth and differentiation of muscle at the transcriptional level.

CLC Number:

S823.2

YANG Tao, XU Houqiang, CHEN Wei, ZHOU Di, WANG Yuanyuan, ZHU Xiaofeng. Study on Interaction between MSTN Promoter and MEF2C Transcription Factor in Cattle[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(8): 1567-1575.

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References

[1] MCPHERRON A C,LAWLER A M,LEE S J.Regulation of skeletal muscle mass in mice by a new TGF-p superfamily member[J].Nature,1997,387(6628):83-90.
[2] SUNDARESAN N R,SAXENA V K,SINGH R,et al.Expression profile of myostatin mRNA during the embryonic organogenesis of domestic chicken (Gallus gallus domesticus)[J].Res Vet Sci,2008,85(1):86-91.
[3] KAMBADUR R,SHARMA M,SMITH T P L,et al.Mutations in myostatin (GDF8) in double-muscled Belgian Blue and Piedmontese cattle[J].Genome Res,1997,7(9):910-915.
[4] MOSHER D S,QUIGNON P,BUSTAMANTE C D,et al.A mutation in the myostatin gene increases muscle mass and enhances racing performance in heterozygote dogs[J].PLoS Genet,2007,3(5):e79.
[5] MOSLER S,RELIZANI K,MOUISEL E,et al.Combinatory effects of siRNA-induced myostatin inhibition and exercise on skeletal muscle homeostasis and body composition[J].Physiol Rep,2014,2(3):e00262.
[6] ZHU X L,HADHAZY M,WEHLING M,et al.Dominant negative myostatin produces hypertrophy without hyperplasia in muscle[J].FEBS Lett,2000,474(1):71-75.
[7] YANG J Z,RATOVITSKI T,BRADY J P,et al.Expression of myostatin pro domain results in muscular transgenic mice[J].Mol Reprod Dev,2001,60(3):351-361.
[8] LEE S J,MCPHERRON A C.Regulation of myostatin activity and muscle growth[J].Proc Natl Acad Sci U S A,2001,98(16):9306-9311.
[9] 王红娜,孙洪新,张英杰,等.干扰MSTN对绵羊成肌细胞增殖分化及相关基因表达的影响[J].畜牧兽医学报,2018,49(1):46-54.
WANG H N,SUN H X,ZHANG Y J,et al.Effects of interfering MSTN on proliferation and differentiation of sheep myoblasts and expression of related genes[J].Acta Veterinaria et Zootechnica Sinica,2018,49(1):46-54.(in Chinese)
[10] FICKETT J W.Quantitative discrimination of MEF2 sites[J].Mol Cell Biol,1996,16(1):437-441.
[11] BOADO R J,PARDRIDGE W M,VINTERS H V,et al.Differential expression of arachidonate 5-lipoxygenase transcripts in human brain tumors:evidence for the expression of a multitranscript family[J].Proc Natl Acad Sci U S A,1992,89(19):9044-9048.
[12] POTTHOFF M J,ARNOLD M A,MCANALLY J,et al.Regulation of skeletal muscle sarcomere integrity and postnatal muscle function by Mef2c[J].Mol Cell Biol,2007,27(23):8143-8151.
[13] NAYAF F J,WU C,RICHARDSON J A,et al.Transcriptional activity of MEF2 during mouse embryogenesis monitored with a MEF2-dependent transgene[J].Development,1999,126(10):2045-2052.
[14] DU R,AN X R,CHEN Y F,et al.Some motifs were important for myostatin transcriptional regulation in sheep (Ovis aries)[J].J Biochem Mol Biol,2007,40(4):547-553.
[15] JUSZCZUK-KUBIAK E,FLISIKOWSKI K,WICINSKA K.Nucleotide sequence and variations of the bovine myocyte enhancer factor 2C (MEF2C) gene promoter in Bos taurus cattle[J].Mol Biol Rep,2011,38(2):1269-1276.
[16] 罗婷,刘军,周云飞,等.Myostatin基因敲除对哺乳期幼鼠骨骼肌生长的影响[J].第三军医大学学报,2017,39(5):401-406.
LUO T,LIU J,ZHOU Y F,et al.Effect of Myostatin knockout on skeletal muscle growth in mice[J].Journal of Third Military Medical University,2017,39(5):401-406.(in Chinese)
[17] SPILLER M P,KAMBADUR R,JEANPLONG F,et al.The myostatin gene is a downstream target gene of basic helix-loop-helix transcription factor MyoD[J].Mol Cell Biol,2002,22(20):7066-7082.
[18] 李佳,邓捷,张军林,等.肌肉增强子因子2对猪肌肉生长抑制素启动子活性的调节[J].生物工程学报,2012,28(8):918-926.
LI J,DENG J,ZHANG J L,et al.Regulation of myostatin promoter activity by myocyte enhancer factor 2[J].Chinese Journal of Biotechnology,2012,28(8):918-926.(in Chinese)
[19] BRYANTSEV A L,BAKER P W,LOVATO T A L,et al.Differential requirements for Myocyte Enhancer Factor-2 during adult myogenesis in Drosophila[J].Dev Biol,2012,361(2):191-207.
[20] EKIYAMA Y,SUZUKI H,TSUKAHARA T.Functional gene expression analysis of tissue-specific isoforms of Mef2c[J].Cell Mol Neurobiol,2012,32(1):129-139.
[21] POTTHOFF M J,OLSON E N.MEF2:a central regulator of diverse developmental programs[J].Development,2007,134(23):4131-4140.
[22] CHEN X,GAO B,PONNUSAMY M,et al.MEF2 signaling and human diseases[J].Oncotarget,2017,8(67):112152-112165.
[23] SCHIAFFINO S,DYAR K A,CALABRIA E.Skeletal muscle mass is controlled by the MRF4-MEF2 axis[J].Curr Opin Clin Nutr Metab Care,2018,21(3):164-167.
[24] JUNG S Y,KO Y G.TRIM72,a novel negative feedback regulator of myogenesis,is transcriptionally activated by the synergism of MyoD (or myogenin) and MEF2[J].Biochem Biophys Res Commun,2010,396(2):238-245.
[25] 夏丹.关岭牛MSTN基因启动子相关转录因子的筛选、克隆及表达纯化[D].贵阳:贵州大学,2016.
XIA D.The screening,cloning,expression and purification of related transcription factors of MSTN gene promoter in Guanling cattle[D].Guiyang:Guizhou University,2016.(in Chinese)
[26] 吴文武.SRF和MEF2及其DNA结合位点保守和演化研究[D].杨凌:西北农林科技大学,2012.
WU W W.Conservation and evolution of SRF and MEF2 with their cognate DNA binding sites[D].Yangling:Northwest A&F University,2012.(in Chinese)
[27] SHAHJAHAN M.Skeletal muscle development in vertebrate animals[J].J Asian Nat Prod Res,2015,1(2):139-148.
[28] WANG B W,CHANG H,KUAN P L,et al.Angiotensin Ⅱ activates myostatin expression in cultured rat neonatal cardiomyocytes via p38 MAP kinase and myocyte enhance factor 2 pathway[J].J Endocrinol,2008,197(1):85-93.
[29] HENNEBRY A,BERRY C,SIRIETT V,et al.Myostatin regulates fiber-type composition of skeletal muscle by regulating MEF2 and MyoD gene expression[J].Am J Physiol Cell Physiol,2009,296(3):C525-C534.

Study on Interaction between MSTN Promoter and MEF2C Transcription Factor in Cattle

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