Differential Regulation of the Promoter Activity of the Mouse UCP2 and UCP3 Genes by MyoD and Myogenin

  • Published : 2007.11.30


UCP2 and UCP3 are members of the uncoupling protein family, which may play roles in energy homeostasis. In order to determine the regulation of the predominant expression of UCP3 in skeletal muscle, the effects of differentiation and myogenic regulatory factors on the promoter activities of the mouse UCP2 and UCP3 genes were studied. Reporter plasmids, containing approximately 3 kb of the 5'-upstream region of the mouse UCP2 and UCP3 genes, were transfected into C2C12 myoblasts, which were then induced to differentiate. Differentiation positively induced the reporter expression about 20-fold via the UCP3 promoter, but by only 2-fold via the UCP2 promoter. C2C12 myoblasts were cotransfected with expression vectors for myogenin and/or MyoD as well as reporter constructs. The simultaneous expression of myogenin and MyoD caused an additional 20-fold increase in the reporter expression via the UCP3 promoter, but only a weak effect via the UCP2 promoter. In L6 myoblasts, only MyoD activated the UCP3 promoter, but in 3T3-L1 cells neither factor activated the UCP3 promoter, indicating that additional cofactors are required, which are present only in C2C12 myoblasts. The expression of UCP2 and UCP3 is differentially regulated during muscle differentiation due to the different responsiveness of their promoter regions to myogenin and MyoD.


  1. Arsenijevic, D., Onuma, H., Pecqueur, C., Raimbault, S., Manning, B., Miroux, B., Couplan, E., Alves-Guerra, M., Goubern, M., Surwit, R., Bouilaud, F., Richard, D., Collins, S. and Ricquier, D. (2000) Disruption of the uncoupling protein 2 gene in mice reveals a role in immunity and reactive oxygen species production. Nature Genet. 26, 435-439.
  2. Boss, O., Samec, S., Paoloni-Giacobina, A., Rossier, C., Dulloo, A., Seydoux, J., Muzzin, P. and Giacobina, J. (1997) Uncoupling protein-3: a new member of the mitochondrial carrier family with tissue-specific expression. FEBS Lett. 408, 39-42.
  3. Chevillotte, E., Rieusset, J., Roques, M., Desage, M. and Vidal, H. (2001) The regulation of uncoupling protein-2 gene expression by w-6 polyunsaturated fatty acids in human skeletal muscle cells involves multiple pathways, including the nuclear receptor peroxisome proliferator-activated receptor b. J. Biol. Chem. 276, 10853-10860.
  4. Dedieu, S., Mazeres, G., Cottin, P. and Brustis, J. (2002) Involvement of myogenic regulator factors during fusion in cell line $C_2C_{12}$. Int. J. Dev. Biol. 46, 235-241
  5. Himms-Hagen, J. and Harper, M. (2001) Physiological role of UCP3 may be export of fatty acids from mitochondria when fatty acid oxidation predominates: an hypothesis. Exp. Biol. Med. 226, 78-84.
  6. Horvath, T., Diano, S., Miyamoto, S., Barry, S., Gatti, S., Alberati, D., Livak, F., Lombardi, A., Moreno, M., Goglia, F., Mor, G., Hamilton, J., Kachinskas, D., Horwitz, B. and Warden, C. (2003) Uncoupling protein 2 and 3 influence obesity and inflammation in transgenic mice. Int. J. Obes. 27, 433-442.
  7. Hwang, C. and Lane, M. D. (1999) Up-regulation of uncoupling protein-3 by fatty acid in C2C12 myotubes. Biochem. Biophys. Res. Commun. 258, 464-469.
  8. Kim, D. (2000) Regulation of expression of uncoupling protein genes, UCP2 and UCP3, in C2C12 muscle cells. MSc Thesis, University of Otago.
  9. Kim, D. (2006) Regulation of UCP2 and UCP3 gene expression. PhD Thesis, University of Otago.
  10. Krauss, S., Zhang, C. and Lowell, B. B. (2005) The mitochondrial uncoupling-protein homologues. Nat. Rev. Mol. Cell Biol. 6, 248-261.
  11. Larkin, S., Mull, E., Miao, W., Pittner, R., Albrandt, K., Moore, C., Young, A., Denaro, M. and Beaumont, K. (1997) Regulation of the third member of the uncoupling protein family, UCP3, by cold and thyroid hormone. Biochem. Biophys. Res. Commun. 240, 222-227.
  12. Marti, A., Larrate, E., Novo, F., Garcia, M. and Martinez, J. (2001) UCP2 muscle gene transfer modifies mitochondrial membrane potential. Int. J. Obes. 25, 68-74.
  13. Matsuda, J., Hosoda, K., Itoh, H., Son, C., Doi, K., Tanaka, T., Fukanaga, Y., Inoue, G., Nishimaru, H., Yoshimasa, Y., Yamori, Y. and Nakao, K. (1997) Cloning of rat uncoupling protein-3 and uncoupling protein-2 cDNAs: their gene expression in rats fed high-fat diet. FEBS Lett. 418, 200-204.
  14. Murre, C., McCaw, P. S., Vaessin, H., Caudy, M., Jan, L. Y., Jan, Y. N., Cabrera, S. V., Buskin, J. N., Hauschka, S. D., Lassar, A. B., Weintraub, H. and Baltimore, D. (1989) Interactions between heterologous helix-loop-helix proteins generate complexes that bind specifically to a common DNA sequence. Cell 58, 537-544.
  15. Nagai, T., Ibata, K., Park, S., Kubota, M., Mikoshiba, K. and Miyawaki, A. (2002) A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological application. Nat. Biotechnol. 20, 87-90.
  16. Nagase, I., Yoshida, S., Canas, X., Irie, Y., Kimura, K., Yoshida, T. and Saito, M. (1999) Up-regulation of uncoupling protein 3 by thyroid hormone, peroxisome proliferator-activated receptor ligands and 9-cis retinoic acid in L6 myotubes. FEBS Lett. 461, 319-322.
  17. Pedraza, N., Rosell, M., Villarroya, J., Iglesias, R., Gonzalez, F. J., Solanes, G. and Villarroya, F. (2006) Development and tissuespecific involvement of peroxisome proliferator-activated receptor-a in the control of mouse uncoupling protein-3 gene expression. Endocrinology 147, 4695-4704.
  18. Schrauwen, P., Hoeks, J., Schaart, G., Kornips, E., Binas, B., Van De Vusse, G., Van Bilsen, M., Luiken, J., Coort, S., Glatz, J., Saris, W. and Hesselink, M. (2003) Uncoupling protein 3 as a mitochondrial fatty acid anion exporter. FASEB J. 17, 2272-2274.
  19. Shimokawa, T., Kato, M., Ezaki, O. and Hashimoto, S. (1998) Transcriptional regulation of muscle-specific genes during myoblast differentiation. Biochem. Biophys. Res. Commun. 246, 287-292.
  20. Solanes, G., Pedraza, N., Inglesias, R., Giralt, M. and Villarroya, F. (2000) The human uncoupling protein-3 gene promoter requires MyoD and is induced by retinoic acid in muscle cells. FASEB J. 14, 2141-2143.
  21. Solanes, G., Pedraza, N., Iglesias, R., Giralt, M. and Villarroya, F. (2003) Functional relationship between MyoD and peroxisome proliferator-activated receptor dependent regulatory pathways in the control of the human uncoupling protein-3 transcription. Mol. Endocrinol. 17, 1944-1958.
  22. Solanes, G., Pedraza, N., Calvo, V., Vidal-Puig, A., Lowell, B. B. and Villarroya, F. (2005) Thyroid hormones directly activate the expression of the human and mouse uncoupling protein-3 genes through a thyroid response element in the proximal promoter region. Biochem. J. 386, 505-513.
  23. Son, C., Hosoda, K., Matsuda, J., Fujikura, J., Yonemitsu, S., Iwakura, H., Masuzaki, H., Ogawa, Y., Hayashi, T., Itoh, H., Nishimura, H., Inoue, G., Yoshimasa, Y., Yamori, Y. and Nakao, K. (2001) Up-regulation of uncoupling protein 3 gene expression by fatty acids and agonists for PPARs in L6 myotubes. Endocrinology 142, 4189-4194.
  24. Thompson, M. P. and Kim, D. (2004) Links between fatty acids and expression of UCP2 and UCP3 mRNAs. FEBS Lett. 568, 4-9.
  25. Vidal-Puig, A., Solanes, G., Grujic, D., Flier, J. and Lowell, B. (1997) UCP3: An uncoupling protein homologue expressed preferentially and abundantly in skeletal muscle and brown adipose tissue. Biochem. Biophys. Res. Commun. 235, 79-82.
  26. Vidal-Puig, A., Grujic, D., Zhang, C., Hagen, T., Boss, O., Ido, Y., Szczepanik, A., Wade, J., Mootha, V., Cortright, R., Muoio, D. and Lowell, B. (2000) Energy metabolism in uncoupling protein 3 gene knockout mice. J. Biol. Chem. 275, 16258-16266.
  27. Weigle, D., Selfridge, L., Schwartz, M., Seeley, R., Cummings, D., Havel, P., Kuijper, J. and BeltrandelRio, H. (1998) Elevated free fatty acids induce uncoupling protein 3 expression in muscle. Diabetes 47, 298-302.
  28. Weintraub, H. (1993) The MyoD family and myogenesis: redundancy, networks and thresholds. Cell 75, 1241-1244.
  29. Wright, W., Sassoon, D. and Lin, V. (1989) Myogenin, a factor regulating myogenesis, has a domain homologous to MyoD. Cell 56, 607-617.
  30. Yoshitomi, H., Yamazaki, K. and Tanaka, I. (1998) Cloning of mouse uncoupling protein 3 cDNA and 5'-flanking region, and its genetic map. Gene 215, 77-84.
  31. Yoshitomi, H., Yamazaki, K. and Tanaka, I. (1999) Mechanism of ubiquitous expression of mouse uncoupling protein 2 mRNA: control by cis-acting DNA element in 5'-flanking region. Biochem. J. 340, 397-404.

Cited by

  1. Leishmania donovani activates uncoupling protein 2 transcription to suppress mitochondrial oxidative burst through differential modulation of SREBP2, Sp1 and USF1 transcription factors vol.48, 2014,
  2. Single nucleotide polymorphisms linked to mitochondrial uncoupling protein genes UCP2 and UCP3 affect mitochondrial metabolism and healthy aging in female nonagenarians vol.17, pp.4, 2016,
  3. Overexpression of NUDT7, a candidate quantitative trait locus for pork color, downregulates heme biosynthesis in L6 myoblasts vol.86, pp.3, 2010,
  4. Serine/threonine protein kinase 25 (STK25): a novel negative regulator of lipid and glucose metabolism in rodent and human skeletal muscle vol.55, pp.6, 2012,
  5. Functional and Activity Analysis of Cattle UCP3 Promoter with MRFs-Related Factors vol.17, pp.5, 2016,