牛MYOZ2基因启动子克隆及活性分析

doi:10.11843/j.issn.0366-6964.2016.04.003

Abstract

Abstract:

The Myozenin2 (MYOZ2) gene promoter of bovine was cloned and sequenced to analyze the active region，which will provide theoretical basis for function determination and expression regulation of MYOZ2 gene.Applying 5′-rapid amplification of cDNA end analysis (RACE)，we determined the transcription initiation site of MYOZ2 gene.Using genome DNA as the template，the promoter of MYOZ2 gene was obtained by PCR.Online software was used to analyze putative transcription factor binding sites.Primer were redesigned according to the results of online software analysis，seven promoter fragments in different length were obtained by unidirectional deletion and cloned into luciferase reporter gene expression vectors.Then，the vectors were transfected into C2C12 cells，their expression activity were measured using the dual reporter assay.The promoter sequence of 2 065 bp of MYOZ2 gene in bovine was obtained and the transcription initiation site of MYOZ2 gene was determined.Luciferase assays indicated that the transcriptional activity of pMYOZ2-84/+125 was significantly higher than pGL3-Basic (P＜0.01)，and the transcriptional activity of pMYOZ2-683/+125 also was significantly higher than pMYOZ2-263/+125 (P＜0.01).The core functional promoter of bovine MYOZ2 was located in the region -84/+125 relative to the TSS，and the transcriptional factors such as MEF2，SRF，MyoD and YY1 might be involved in the transcriptional regulation of MYOZ2 gene.

Key words: bovine, MYOZ2 promoter, gene cloning, activity analysis, 5′RACE

CLC Number:

WANG Ming-ming，ZHAO Zhi-dong，LI An-ning，ZHANG Ya-ran，DUAN Mei-yan，LI Shi-jun，WU Sen，WANG Xiao-yu，ZAN Lin-sen. Cloning and Activity Analysis of Bovine MYOZ2 Gene Promoter[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(4): 652-660.

References

［1］刘居鹏，单安山，彭济昌.猪肌纤维发育及其与肉质的关系［J］.中国饲料，2006(16)：11-14.
LIU J P，SHAN A S，PENG J C.Development of pig muscle fiber and the relationship with the meat quality［J］.China Feed，2006(16)：11-14.(in Chinese)
［2］陈代文，张克英，胡祖禹.猪肉品质特征的形成原理［J］.四川农业大学学报，2002，20(1)：60-66.
CHEN D W，ZHANG K Y，HU Z Y.The mechanism for developing pork quality traits［J］.Journal of Sichuan Agricultural University，2002，20(1)：60-66.(in Chinese)
［3］FREY N，RICHARDSON J A，OLSON E N.Calsarcins，a novel family of sarcomeric calcineurin-binding proteins［J］. Proc Natl Acad Sci U S A，2000，97(26)：14632-14637.
［4］OSIO A，TAN L，CHEN S N，et al.Myozenin 2 is a novel gene for human hypertrophic cardiomyopathy［J］.Circ Res，2007，100(6)：766-768.
［5］FREY N，BARRIENTOS T，SHELTON J M，et al.Mice lacking calsarcin-1 are sensitized to calcineurin signaling and show accelerated cardiomyopathy in response to pathological biomechanical stress［J］.Nat Med，2004，10(12)：1336-1343.
［6］WANG H，YANG S，YANG E，et al.NF-kappaB mediates the transcription of mouse calsarcin-1 gene，but not calsarcin-2，in C2C12 cells［J］.BMC Mol Biol，2007，8：19.
［7］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.
DUAN M Y，ZHAO Z D，LI A N，et al.Analysis of bovine ATP5B gene promoter and tissue expression［J］.Acta Veterinaria et Zootechnica Sinica，2015，46(3)：382-387.(in Chinese)
［8］CENIK B K，GARG A，MCANALLY J R，et al.Severe myopathy in mice lacking the MEF2/SRF-dependent gene leiomodin-3［J］. J Clin Invest，2015，125(4)：1569-1578.
［9］HERNDON C A，ANKENBRUCK N，LESTER B，et al.Neuregulin1 signaling targets SRF and CREB and activates the muscle spindle-specific gene Egr3 through a composite SRF-CREB-binding site［J］. Exp Cell Res，2013，319(5)：718-730.
［10］COLLARD L，HERLEDAN G，PINCINI A，et al.Nuclear actin and myocardin-related transcription factors control disuse muscle atrophy through regulation of Srf activity［J］.J Cell Sci，2014，127( Pt 24)：5157-5163.
［11］GUERCI A，LAHOUTE C，HÉBRARD S，et al.Srf-dependent paracrine signals produced by myofibers control satellite cell-mediated skeletal muscle hypertrophy［J］.Cell Metab，2012，15(1)：25-37.
［12］JANG Y N，BAIK E J.JAK-STAT pathway and myogenic differentiation［J］.JAKSTAT，2013，2(2)：e23282.
［13］SINGH K，CASSANO M，PLANET E，et al.A KAP1 phosphorylation switch controls MyoD function during skeletal muscle differentiation［J］. Genes Dev，2015，29(5)：513-525.
［14］HWANG J，LEE S J，YOO M，et al.Kazinol-P from Broussonetia kazinoki enhances skeletal muscle differentiation via p38MAPK and MyoD［J］. Biochem Biophys Res Commun，2015，456(1)：471-475.
［15］EKMARK M，RANA Z A，STEWART G，et al.De-phosphorylation of MyoD is linking nerve-evoked activity to fast myosin heavy chain expression in rodent adult skeletal muscle［J］.J Physiol，2007，584(Pt 2)：637-650.
［16］BLUM R.Activation of muscle enhancers by MyoD and epigenetic modifiers［J］.J Cell Biochem，2014，115(11)：1855-1867.
［17］ESTRELLA N L，DESJARDINS C A，NOCCO S E，et al.MEF2 transcription factors regulate distinct gene programs in mammalian skeletal muscle differentiation［J］.J Biol Chem，2015，290(2)：1256-1268.
［18］CAINE C，KASHEROV P，SILBER J，et al.Mef2 interacts with the Notch pathway during adult muscle development in Drosophila melanogaster［J］.PLoS One，2014，9(9)：e108149.
［19］WALES S，HASHEMI S，BLAIS A，et al.Global MEF2 target gene analysis in cardiac and skeletal muscle reveals novel regulation of DUSP6 by p38MAPK-MEF2 signaling［J］.Nucleic Acids Res，2014，42(18)：11349-11362.
［20］VARGAS M A，TIRNAUER J S，GLIDDEN N，et al.Myocyte enhancer factor 2 (MEF2) tethering to muscle selective A-kinase anchoring protein (mAKAP) is necessary for myogenic differentiation［J］.Cell Signal，2012，24(8)：1496-1503.
［21］LEE T C，ZHANG Y，SCHWARTZ R J.Bifunctional transcriptional properties of YY1 in regulating muscle actin and c-myc gene expression during myogenesis［J］.Oncogene，1994，9(4)：1047-1052.
［22］GALVAGNI F，CARTOCCI E，OLIVIERO S.The dystrophin promoter is negatively regulated by YY1 in undifferentiated muscle cells［J］.J Biol Chem，1998，273(50)：33708-33713.
［23］WALOWITZ J L，BRADLEY M E，CHEN S，et al.Proteolytic regulation of the zinc finger transcription factor YY1，a repressor of muscle-restricted gene expression［J］.J Biol Chem，1998，273(12)：6656-6661.
［24］WANG H，YANG S，YANG E，et al.NF-kappaB mediates the transcription of mouse calsarcin-1 gene，but not calsarcin-2，in C2C12 cells［J］.BMC Mol Biol，2007，8：19.

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Cloning and Activity Analysis of Bovine MYOZ2 Gene Promoter

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