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Functional analysis of pig myostatin gene promoter with some adipogenesis- and myogenesis-related factors

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Abstract

Myostatin (MSTN) is primarily expressed in muscle and plays an important role in muscle and fat development in pigs. However, there is little information about the regulation of pig MSTN. In order to elucidate whether pig MSTN could be regulated by muscle- and fat-related factors, the porcine MSTN promoter was amplified and cloned into pGL3-basic vector, and transfected into cells to analyze the transcriptional activity of promoter with muscle- and fat-related factors through Dual-luciferase reporter assays. 5′-deletion expression showed that there was a negative-regulatory region located between nucleotides −1519 and −1236 bp, and there were some positive-regulatory regions located between −1236 and −568 bp. The longest fragment (1.7 kb) was cotransfected with muscle-related transcription factor myogenic differentiation 1 (MyoD), resulting in promoter transcriptional activity upregulation. The fragment was treated by the adipogenic agents (DIM) including dexamethasone, insulin, and isobutyl-1-methylxanthine (IBMX). We found that MSTN promoter transcriptional activity can be regulated by IBMX, but not by DIM. CCAAT/enhancer binding protein (C/EBP) α and C/EBPβ, two proteins which are induced by DIM during adipogenesis were cotransfected with the 1.7-kb fragment, respectively, resulting in promoter transcriptional activity downregulation. Treating the fragment with rosiglitazone which induce the expression of peroxisome proliferator-activated receptor γ (PPARγ), resulting in promoter transcriptional activity upregulation. Cotransfection experiments confirmed this result. Taken together, we showed that porcine MSTN could be upregulated by IBMX, MyoD, and PPARγ but downregulated by C/EBPα and C/EBPβ.

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References

  1. Langley B, Thomas M, Bishop A, Sharma M, Gilmour S, Kambadur R (2002) Myostatin inhibits myoblast differentiation by down-regulating MyoD expression. J Biol Chem 277(51):49831–49840. doi:10.1074/jbc.M204291200M204291200

    Article  PubMed  CAS  Google Scholar 

  2. McPherron AC, Lee SJ (1997) Double muscling in cattle due to mutations in the myostatin gene. Proc Natl Acad Sci USA 94(23):12457–12461

    Article  PubMed  CAS  Google Scholar 

  3. Thomas M, Langley B, Berry C, Sharma M, Kirk S, Bass J, Kambadur R (2000) Myostatin, a negative regulator of muscle growth, functions by inhibiting myoblast proliferation. J Biol Chem 275(51):40235–40243. doi:10.1074/jbc.M004356200M004356200

    Article  PubMed  CAS  Google Scholar 

  4. Kambadur R, Sharma M, Smith TPL, Bass JJ (1997) Mutations in myostatin (GDF8) in double-muscled Belgian Blue and Piedmontese cattle. Genome Res 7(9):910–916

    PubMed  CAS  Google Scholar 

  5. Guo T, Jou W, Chanturiya T, Portas J, Gavrilova O, McPherron AC (2009) Myostatin inhibition in muscle, but not adipose tissue, decreases fat mass and improves insulin sensitivity. PLoS One 4(3):e4937. doi:10.1371/journal.pone.0004937

    Article  PubMed  Google Scholar 

  6. Artaza JN, Bhasin S, Magee TR, Reisz-Porszasz S, Shen R, Groome NP, Meerasahib MF, Gonzalez-Cadavid NF (2005) Myostatin inhibits myogenesis and promotes adipogenesis in C3H 10T(1/2) mesenchymal multipotent cells. Endocrinology 146(8):3547–3557. doi:10.1210/en.2005-0362

    Article  PubMed  CAS  Google Scholar 

  7. Kim HS, Liang L, Dean RG, Hausman DB, Hartzell DL, Baile CA (2001) Inhibition of preadipocyte differentiation by myostatin treatment in 3T3–L1 cultures. Biochem Biophys Res Commun 281(4):902–906. doi:10.1006/bbrc.2001.4435S0006-291X(01)94435-3

    Article  PubMed  CAS  Google Scholar 

  8. Patruno M, Caliaro F, Maccatrozzo L, Sacchetto R, Martinello T, Toniolo L, Reggiani C, Mascarello F (2008) Myostatin shows a specific expression pattern in pig skeletal and extraocular muscles during pre- and post-natal growth. Differentiation 76(2):168–181

    Article  PubMed  CAS  Google Scholar 

  9. Yang JZ, Schulman L, Pursel VG, Solomon MB, Mitchell A, Zhao BP, Li ZC, Wall RJ (2010) Expression of porcine myostatin prodomain genomic sequence leads to a decrease in muscle growth, but significant intramuscular fat accretion in transgenic pigs. Transgenic Res 19(1):149

    Google Scholar 

  10. Guimaraes SEF, Stahl CH, Lonergan SM, Geiger B, Rothschild MF (2007) Myostatin promoter analysis and expression pattern in pigs. Livestock Sci 112(1–2):143–150

    Article  Google Scholar 

  11. Stinckens A, Luyten T, Bijttebier J, Van den Maagdenberg K, Dieltiens D, Janssens S, De Smet S, Georges M, Buys N (2008) Characterization of the complete porcine MSTN gene and expression levels in pig breeds differing in muscularity. Anim Genet 39(6):586–596

    Article  PubMed  CAS  Google Scholar 

  12. Spiller MP, Kambadur R, Jeanplong F, Thomas M, Martyn JK, Bass JJ, Sharma M (2002) The myostatin gene is a downstream target gene of basic helix-loop-helix transcription factor MyoD. Mol Cell Biol 22(20):7066–7082

    Article  PubMed  CAS  Google Scholar 

  13. Chen WW, Wu WF, Zhao J, Yu CX, Liu WW, Jiang AL, Zhang JY (2009) Molecular cloning and preliminary analysis of the human alpha-methylacyl-CoA racemase promoter. Mol Biol Rep 36(3):423–430

    Article  PubMed  CAS  Google Scholar 

  14. Funkenstein B, Balas V, Rebhan Y, Pliatner A (2009) Characterization and functional analysis of the 5’ flanking region of Sparus aurata myostatin-1 gene. Comp Biochem Physiol A 153(1):55–62

    Article  Google Scholar 

  15. Yu Z, Li Y, Meng Q, Yuan J, Zhao Z, Li W, Hu X, Yan B, Fan B, Yu S, Li N (2005) Comparative analysis of the pig BAC sequence involved in the regulation of myostatin gene. Sci China C Life Sci 48(2):168–180

    Google Scholar 

  16. Ma K, Mallidis C, Artaza J, Taylor W, Gonzalez-Cadavid N, Bhasin S (2001) Characterization of 5′-regulatory region of human myostatin gene: regulation by dexamethasone in vitro. Am J Physiol Endocrinol Metab 281:E1128–E1136

    PubMed  CAS  Google Scholar 

  17. Du R, An XR, Chen YF, Qin J (2007) Some motifs were important for myostatin transcriptional regulation in sheep (Ovis aries). J Biochem Mol Biol 40(4):547–553

    Article  PubMed  CAS  Google Scholar 

  18. Allen DL, Du M (2008) Comparative functional analysis of the cow and mouse myostatin genes reveals novel regulatory elements in their upstream promoter regions. Comp Biochem Physiol 150(4):432–439

    Article  Google Scholar 

  19. Hennebry A, Berry C, Siriett V, O’Callaghan P, Chau L, Watson T, Sharma M, Kambadur R (2009) Myostatin regulates fiber-type composition of skeletal muscle by regulating MEF2 and MyoD gene expression. Am J Physiol Cell Physiol 296(3):C525–C534. doi:10.1152/ajpcell.00259.2007

    Article  PubMed  CAS  Google Scholar 

  20. Feldman BJ, Streeper RS, Farese RV Jr, Yamamoto KR (2006) Myostatin modulates adipogenesis to generate adipocytes with favorable metabolic effects. Proc Natl Acad Sci USA 103(42):15675–15680. doi:10.1073/pnas.0607501103

    Google Scholar 

  21. Thonpho A, Sereeruk C, Rojvirat P, Jitrapakdee S (2010) Identification of the cyclic AMP responsive element (CRE) that mediates transcriptional regulation of the pyruvate carboxylase gene in HepG2 cells. Biochem Biophys Res Commun 393(4):714–719

    Article  PubMed  CAS  Google Scholar 

  22. Ehrmann J, Vavrusova N, Collan Y, Kolar Z (2002) Peroxisome proliferator-activated receptors (PPARs) in health and disease. Biomed Papers 146(2):11–14

    CAS  Google Scholar 

  23. Allen DL, Cleary AS, Hanson AM, Lindsay SF, Reed JM (2010) CAAT/enhancer binding protein-delta expression is increased in fast skeletal muscle by food deprivation and regulates myostatin transcription in vitro. Am J Physiol Regul Integr Comp Physiol 299(6):R1592–R1601

    Article  PubMed  CAS  Google Scholar 

  24. White UA, Stephens JM (2010) Transcriptional factors that promote formation of white adipose tissue. Mol Cell Endocrinol 318(1–2):10–14

    Article  PubMed  CAS  Google Scholar 

  25. Chawla A, Schwarz EJ, Dimaculangan DD, Lazar MA (1994) Peroxisome proliferator-activated receptor (PPAR) gamma: adipose-predominant expression and induction early in adipocyte differentiation. Endocrinology 135(2):798–800

    Article  PubMed  CAS  Google Scholar 

  26. Porse BT, Pedersen T, Xu X, Lindberg B, Wewer UM, Friis-Hansen L, Nerlov C (2001) E2F repression by C/EBP [alpha] is required for adipogenesis and granulopoiesis in vivo. Cell 107(2):247–258

    Article  PubMed  CAS  Google Scholar 

  27. Stein B, Yang MX (1995) Repression of the interleukin-6 promoter by estrogen receptor is mediated by NF-kappa B and C/EBP beta. Mol Cell Biol 15(9):4971–4979

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by the major projects of Genetically Modified Animals (2009ZX08009-161B), the creative team project of Chinese education ministry (IRT-0831) and National Natural Science Foundation of China (30871779).

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Authors and Affiliations

  1. Agricultural Ministry Key Laboratory of Swine Breeding and Genetics and Key Laboratory of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, 430070, Hubei, People’s Republic of China

    Bing Deng, Yi Ding & Siwen Jiang

  2. Institute of Animal Science and Veterinary Medicine of Wuhan, Wuhan Academy of Agricultural Science & Technology, Wuhan, 430065, Hubei, People’s Republic of China

    Bing Deng, Qishuang Gao, Haijun Huang, Zhiping Ran & Yunguo Qian

  3. School of Mathematics and Statistics, Huazhong University of Science and Technology, Wuhan, 430074, People’s Republic of China

    Jianghui Wen

  4. Department of Animal Nutrition and Feed Science, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, 430070, Hubei, People’s Republic of China

    Jian Peng

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  1. Bing Deng
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  2. Jianghui Wen
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  3. Yi Ding
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  7. Yunguo Qian
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  9. Siwen Jiang
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Corresponding authors

Correspondence to Jian Peng or Siwen Jiang.

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Siwen Jiang and Jian Peng contributed equally to this work.

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Deng, B., Wen, J., Ding, Y. et al. Functional analysis of pig myostatin gene promoter with some adipogenesis- and myogenesis-related factors. Mol Cell Biochem 363, 291–299 (2012). https://doi.org/10.1007/s11010-011-1181-y

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  • DOI: https://doi.org/10.1007/s11010-011-1181-y

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