The uncoupling protein family is a mitochondrial anion carrier family. It plays an important role in the biological traits of animal body weight, basal metabolic rate and energy conversion. Using PCR and PCR-SSCP, we scanned the porcine uncoupling protein 2 gene (UCP2) and uncoupling protein 3 gene (UCP3) and found seven deletion sites, three in UCP2 and four in UCP3. The deletions in 15 pig breeds showed that deletion influenced weight. The genotype compounding of seven deletion sites in 15 pig breeds had significant effects on performance traits of the pig, such as body weight. We predicted the potential protein factor binding sites using the transcription factor analysis tool TFSearch version 1.3 online. Two deletions (1830 nt and 3219 nt) in UCP3 were found to change the transcriptional factor sites. The 16 bp deletion in 1830 nt added a SP1 site and a 6 bp deletion in 3219 nt removed two MZF1 sites. Seven deletion polymorphisms were covered in introns of linkage genes of UCP2 and UCP3, showing that UCPs have conservation and genetic reliability.
Similar content being viewed by others
REFERENCES
Bassam, B. J., Caetano-Anolles, G., and Gresshoff, P. M. (1991). Fast and sensitive silver staining of DNA in polyacrylamide gels. Anal. Biochem. 196:80–83.
Black, A. R., Jensen, D., Lin, S-Y, and Azizkhan, J. C. (1999). Growth/cell cycle regulation of Sp1 phosphorylation. J. Biol. Chem. 274:1207–1215.
Damon, M., Vincent, A., Lombardi, A., and Herpin, P. (2000). First evidence of uncoupling protein-2 (UCP-2) and -3 (UCP-3) gene expression in piglet skeletal muscle and adipose tissue. Gene 246:133–141.
Enerback, S., Jacobsson, A., Simpson, E. M., Guerra, C., Yamashita, H., Harper, M. E., and Kozak, L. P. (1997). Mice lacking mitochondrial uncoupling protein are cold-sensitive but not obese. Nature 387:90–94.
Esterbauer, H., Schneitler, C., Oberkofler, H., Ebenbichler, C., Paulweber, B., Sandhofer, F., Ladurner, G., Hell, E., Strosberg, A. D., Patsch, J. R., Krempler, F., and Patsch, W. (2001). A common polymorphism in the promoter of UCP2 is associated with decreased risk of obesity in middle-aged humans. Nat. Genet. 28:178–183.
Fang, M., Liu, C., Zhao, X., Li, N., and Wu, C. (2002a). Sequences and polymorphisms of exons 3 and 4 in porcine UCP2 gene. Progress Nat Sci 12:551–552.
Fang, M., Zhao, X., Li, N., and Wu, C. (2002b). Genetic analysis on 3′-terminal flanking region of uncoupling protein 3 in different pig breeds. Chinese Sci. Bull. 47:1541–1543.
Hir, H. L., Nott, A., and Moore, M. J. (2003). How introns influence and enhance eukaryotic gene expression. Trends Biochem. Sci. 28:215–220.
Hromas, R., Collins, S. J., Hickstein, D., Raskind, W., Deaven, L. L., O'Hara, P., Hagen, F. S., and Kaushansky, K. (1991). A retinoic acid-responsive human zinc finger gene, MZF-1, preferentially expressed in myeloid cells. J. Biol. Chem. 266:14183–14187.
Katsumata, M., Matsumoto, M., Kawakami, S., and Kaji, Y. (2004). Effect of heat exposure on uncoupling protein-3 mRNA abundance in porcine skeletal muscle. J. Anim. Sci. 82:3493–3499.
Krempler, F., Esterbauer, H., Weitgasser, R., Ebenbichler, C., Patsch, J. R., Miller, K., Xie, M., Linnemayr, V., Oberkofler, H., and Patsch, W. (2002). A functional polymorphism in the promoter of UCP2 enhances obesity risk but reduces type 2 diabetes risk in obese middle-aged humans. Diabetes 51:3331–3335.
Li, H., Li, Y., Zhao, X., Li, N., and Wu, C. (2005). Structure and nucleotide polymorphisms in pig uncoupling protein 2 and 3 genes. Anim. Biotechnol. 16:209–220.
Long, M., de Souza, S. J., and Gilbert, W. (1995). Evolution of the intron-exon structure of eukaryotic genes. Curr. Opin. Genet. 5:774–778.
Mattick, J. S. (1994). Intron: Evolution and function. Curr. Opin. Genet. Dev. 4:823–831.
Moore, M. J. (2003). A quantitative analysis of intron effects on mammalian gene expression. RNA 9:607–617.
Morris, J. F., Rauscher, F. J. III, Davis, B., Klemsz, M., Xu, D., Tenen, D., and Hromas, R. (1995). The myeloid zinc finger gene, MZF-1, regulates the CD34 promoter in vitro. Blood 86:3640–3647.
Mostyn, A., Litten, J. C., Perkins, K. S., Alves-Guerra, M. C., Pecqueur, C., Miroux, B., Symonds, M. E., and Clarke, L. (2004). Influence of genotype on the differential ontogeny of uncoupling protein 2 and 3 in subcutaneous adipose tissue and muscle in neonatal pigs. J. Endocrinol. 183:121–131.
Mostyn, A., Litten, J. C., Perkins, K. S., Euden P. J., Corson, A. M., Symonds, M. E., and Clarke, L. (2005). Influence of size at birth on the endocrine profiles and expression of uncoupling proteins in subcutaneous adipose tissue, lung, and muscle of neonatal pigs. Am. J. Physiol. Regul. Integr. Comp. Physiol. 288:1536–1542.
Otabe, S., Clement, K., Rich, N., Warden, C., Pecqueur, C., Neverova, M., Raimbault, S., Guy-Grand, B., Basdevant, A., Ricquier, D., Froguel, P., and Vasseur, F. (1998). Mutation screening of the human UCP 2 gene in normoglycemic and NIDDM morbidly obese patients: lack of association between new UCP 2 polymorphisms and obesity in French Caucasians. Diabetes 47:840–842.
Pei, D. Z. (1996). General situation of Chinese miniature pig. Rural Animal-Production Technologe 8:18.
Schonfeld-Warden, N. A., and Warden, C. H. (2001). Physiological effects of variants in human uncoupling proteins: UCP2 influences body-mass index. Biochem. Soc. Trans. 29:777–784.
Suske, G. (1999). The Sp-family of transcription factors. Gene 238:291–300.
Tu, R. J., Deng, C. Y., and Xiong, Y. Z. (2004). Sequencing analysis on partial translated region of uncoupling protein-3 gene and single nucleotide polymorphism associations with the porcine carcass and meat quality traits. Act. Genet. Sinic. 31:807–812.
Walder, K., Norman, R. A., Hanson, R. L., Schrauwen, P., Neverova, M., Jenkinson, C. P., Easlick, J., Warden, C. H., Pecqueur, C., Raimbault, S., Ricquier, D., Silver, M. H., Shuldiner, A. R., Solanes, G., Lowell, B. B., Chung, W. K., Leibel, R. L., Pratley, R., and Ravussin, E. (1998). Association between uncoupling protein polymorphisms (UCP2-UCP3) and energy metabolism/obesity in Pima Indians. Hum. Mol. Genet. 7:1431–1435.
Werner, P., Neuenschwander, S., and Stranzinger, G. (1999). Characterization of the porcine uncoupling proteins 2 and 3 (UCP2 and UCP3) and their localization to chromosome 9 p by somatic cell hybrids. Anim. Genet. 30:221–224.
Yeung, F., Law, W. K., Yeh, C. H., Westendorf, J. J., Zhang, Y., Wang, R., Kao, C., and Chung, L. W. (2002). Regulation of human osteocalcin promoter in hormone-independent human prostate cancer cells. J. Biol. Chem. 277:2468–2476.
Zawia, N. H., Sharan, R., Brydie, M., Oyama, T., and Crumpton, T. (1998). Sp1 as a target site for metal-induced perturbations of transcriptional regulation of developmental brain gene expression. Dev. Brain Res. 107:291–298.
ACKNOWLEDGMENTS
This work was supported by the National Natural Science Foundation of China (Grant No. 30471235), the National Basic Research Program of China (2006CB102100) and Fok Ying Tung Education Foundation (Grant No. 91028).