Abstract
Leptin is expressed mainly by adipocytes and plays a crucial role in the regulation of energy expenditure, food intake, and adiposity. Using PCR-heteroduplex analysis and sequencing, we investigated a C/G substitution in the promoter region of the bovine leptin gene. Application of the electrophoretic mobility shift assay showed that the C→G transversion decreased the leptin gene promoter binding capacity for nuclear proteins. With real-time PCR and Western blotting, we showed that the leptin expression level was higher in cattle with the CC than with the GG genotype.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Andrews NC, Faller DV (1991) A rapid micropreparation technique for extraction of DNA-binding proteins from limiting numbers of mammalian cells. Nucleic Acids Res 19: 2499
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159
Considine RV, Sihna MK, Heiman ML, Kriauciunas A, Stephens TW, et al. (1996) Serum immunoreactive-leptin concentrations in normal-weight and obese humans. N Engl J Med 334: 292–295
de Vos P, Saladin R, Auwerx J, Staels B (1995) Induction of ob gene expression by corticosteroids is accompanied by body weight loss and reduced food intake. J Biol Chem 270: 15958–15961
Flier JS (1997) Leptin expression and action: new experimental paradigms. Proc Natl Acad Sci U S A 94: 4242–4245
Fukuda H, Iritani N (1999) Transcriptional regulation of leptin gene promoter in rat. FEBS Lett 455: 165–169
Haegeman A, van Zeveren A, Peelman LJ (2000) New mutation in exon 2 of the bovine leptin gene. Anim Genet 31: 79
Kanai N, Fujii T, Saito K, Yokoyama T (1994) Rapid and simple method for preparation of genomic DNA from easily obtained clotted blood. J Clin Pathol 47: 1043–1044
Konfortov BA, Licence VE, Miller JR (1999) Re-sequencing of DNA from a diverse panel of cattle reveals a high level of polymorphism in both intron and exon. Mamm Genome 10: 1142–1145
Lagonigro R, Wiener P, Pilla F, Wooliams JA, Williams JL (2003) A new mutation in the coding region of the bovine leptin gene associated with feed intake. Anim Genet 34: 371–374
Leifers S (2004) Physiology and genetics of leptin in periparturient dairy cows. Ph.D thesis, Wageningen University
Liefers SC, Veerkamp RF, te Pas MF, Delavaud C, Chilliard Y, et al. (2005) Leptin promoter mutations affect leptin levels and performance traits in dairy cows. Anim Genet 36: 111–118
Malewski T, Gajewska M, Żebrowska T, Zwierzchowski L (2002) Differential induction of transcription factors and expression of milk protein genes by prolactin and growth hormone in the mammary gland of rabbits. Growth Horm IGF Res 12: 41–53
Mason MM, Fe Y, Chen H, Quon MJ, Reitam M (1998) Regulation of leptin promoter function by Sp1, C/EBP, and a novel factor. Endocrinology 139: 1013–1022
Pomp D, Zou T, Clutter AC, Barendse W (1997) Rapid communication: mapping of leptin to bovine chromosome 4 by linkage analysis of a PCR-based polymorphism. J Anim Sci 75: 1427
Slieker LJ, Sloop KW, Surface PL, Kriaciunas A, LaQuier F, et al. (1996) Regulation of the expression of ob mRNA and protein by glucocortycoids and cAMP. J Biol Chem 271: 5301–5304
Taniguchi Y, Itoh T, Yamada T, Sasaki Y (2002) Genomic structure and promoter analysis of the bovine leptin gene. IUBMB Life 53: 131–135
Acknowledgments
This study was funded by the State Committee for Scientific Research (KBN) grant 2 P06D 044 26.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Adamowicz, T., Flisikowski, K., Starzyński, R. et al. Mutation in the Sp1 motif of the bovine leptin gene affects its expression. Mamm Genome 17, 77–82 (2006). https://doi.org/10.1007/s00335-005-0068-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00335-005-0068-1