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Molecular Cloning and mRNA Expression of the Bovine Peroxisome Proliperator Receptor Gamma(PPARγ)

한우 PPARγ 유전자의 동정과 mRNA의 발현

  • Jeoung, Y.H. (Department of Animal Science, Insti. of Ag. Sci. and Tech., College of Agriculture & Life Science, Chonnam National University) ;
  • Lee, S.M. (Department of Animal Science, Insti. of Ag. Sci. and Tech., College of Agriculture & Life Science, Chonnam National University) ;
  • Park, H.Y. (Department of Animal Science, Insti. of Ag. Sci. and Tech., College of Agriculture & Life Science, Chonnam National University) ;
  • Yoon, D.H. (National Liverstock Research Institute) ;
  • Choi, J.G. (National Liverstock Research Institute, Daekwanryeong Branch) ;
  • Moon, S.J. (Department of Animal Science, Insti. of Ag. Sci. and Tech., College of Agriculture & Life Science, Chonnam National University) ;
  • Kang, M.J. (Department of Animal Science, Insti. of Ag. Sci. and Tech., College of Agriculture & Life Science, Chonnam National University)
  • 정영희 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 이상미 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 박효영 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 윤두학 (축산연구소) ;
  • 최재관 (축산연구소 대관령지소) ;
  • 문승주 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 강만종 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부)
  • Published : 2004.02.29

Abstract

The peroxisome proliferator-activated receptor $\gamma$(PPAR$\gamma$), a member of the steroid/thyroid nuclear hormone receptor suferfamily of ligand-activated transcription factor, is an important regulator of adipocyte gene expression and differentiation. In this studies, we report the identification, characterization, and expression of a Hanwoo PPAR$\gamma$ gene. The PPAR$\gamma$ cDNA sequence of the Hanwoo show strong conservation with the corresponding sequences reported in other species except of three amino acid sequences. The distribution of PPAR$\gamma$ mRNA in various tissues of Korean cattle aged 12 months were investigated using Northern Blot analysis. The highest expression was detected in adipose tissue, more lower expression was detected in colon, small intestine, kidney, lung, while expression was not detected in brain, heart. PPAR$\gamma$ expression was higher in adipose tissue of Korean cattle when aged 30 months than aged 12 months. These results indicated PPAR$\gamma$, regulator adipocyte gene expression and differentiation, related on adipose differentiation in Korean native cattle(HANWOO).

지방세포분화 과정에서 중추적인 역할을 한다고 보고되어지고 있는 PPAR$\gamma$ 유전자를 cloning 하기 위하여 한우 12개월 지방조직을 이용하여 Total RNA를 추출하고 RT-PCR을 수행하여 1515bp, 505 아미노산 서열을 가진 PPAR$\gamma$ 유전자를 cloning 하였다. bovine을 제외한 4종의 다른 동물과의 아미노산 서열을 비교한 결과 모두 90% 이상의 상동성을 나타내었다. 특히 NCBl에 보고된 Norwegian cattle PPAR$\gamma$ (NCBI Accession No. O18971)의 아미노산 서열과 비교한 결과 99.2%의 상동성을 나타내었으며 아미노산 서열중 81번째 이소루신, 140번째 아르기닌, 157번째 글루탐산, 486번째 리신이 각각 트레오닌, 트립토판, 글리신, 이소루신으로 치환된 상태의 아미노산 서열을 나타내었다. 특히 nuclear hormone receptor의 DNA binding domain 영역에 포함 되는 140번째, 157번째 아미노산의 치환은 앞으로 한우PPAR$\gamma$ 유전자가 조절하는 다른 유전자와의 관계를 밝히는데 매우 중요한 점이 될 것으로 사료되어진다. 한우 PPAR$\gamma$ 의 발현은 지방조직에서 매우 높은 발현을 보였으며, 한우 12개월령과 30개월령 지방조직에서의 PPAR$\gamma$ 발현을 비교 분석하였을 때 12개월령에서보다 30개월령 지방조직에서 PPAR$\gamma$ 발현이 약 6배정도 높았다. 이와 같은 결과는 한우 PPAR$\gamma$ 유전자도 사람과 생쥐에서와 같이 지방분화에 관여하고 있다는 보고와 일치하는 결과라고 사료된다.

Keywords

References

  1. Brun, R. P. and Spiegelman, B. M. 1997. PPAR gamma and the molecular control of adipogenesis. J Endocrinol 155:217.
  2. Brun, R. P., Tontonoz, P., Forman, B. M., Ellis, R, Chen, J., Evans, R. M. and Spiegelman, B. M. 1996. Differential activation of adipogenesis by multiple PPAR isoforms. Genes Dev 10:974.
  3. Chawla, A. and Lazar, M. A. 1994. Peroxisome proliferator and retinoid signaling pathways coregulate preadipocyte phenotype and survival. Proc Natl Acad Sci. V.SA. 91:1786.
  4. Forman, B. M., Tontonoz, P., Chen, 1., Brun, R. P., Spiegelman, B. M. and Evans, R M. 1995. 15-Deoxy-delta 12, 14-prostaglandin J2 is a ligand for the adipocyte determination factor PPAR gamma Cell. 83:803.
  5. Gottlicher, M., Widmark, E., Li, Q. and Gustafsson, J. A. 1992. Fatty acids activate a chimera of the clofibric acid-activated receptor and the glucocorticoid receptor. Proc Natl Acad Sci. V.SA. 89:4653.
  6. Grimaldi, P. A. 2001. The role of PPARs in adipocyte differentiation. Prog. Lipid. Res. 40:269.
  7. Kliewer, S. A., Umesono, K, Mangelsdorf, D. J. and Evans, R M. 1992. Retinoid X receptor interacts with nuclear receptors in retinoic acid, thyroid hormone and vitamin D3 signalling. Nature. 355:444
  8. Kliewer, S. A., Lenhard, J. M., Willson, T. M., Patel, I., Morris, D. C. and Lehmann, J. M. 1995. A prostaglandin J2 metabolite binds peroxisome proliferator-activated receptor gamma and promotes adipocyte differentiation. Cell. 83:813.
  9. Perlmann, T. and Jansson, L. 1995. A novel pathway for vitamin A signaling mediated by RXR heterodimerization with NGFl-B and NURRI. Genes Dev. 9:769.
  10. Sambrook, J. and Russell, D. W. 2001. Molecular Cloning: A Loboratory Manual, 3rd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
  11. Schoonjans, K., Staels, B. and Auwerx, J. 1996. The peroxisome proliferator activated receptors (PPARS) and their effects on lipid metabolism and adipocyte differentiation. Biochim Biophys Acta 1302:93.
  12. Sundvold, H., Brzozowska, A. and Lien, S. 1997. Characterisation of bovine peroxisome proliferatoractivated receptors gamma I and gamma 2: genetic mapping and differential expression of the two isoforms. Biochem Biophys Res Commun. 239:857.
  13. Tontonoz, P., Hu, E., Graves, R. A., Budavari, A. I. and Spiegelman, B. M. 1994. mPPAR gamma 2: tissue-specific regulator of an adipocyte enhancer. Genes Dev. 8:1224.
  14. Tontonoz, P., Hu, E. and Spiegelman, B. M. 1994. Stimulation of adipogenesis in fibroblasts by PPAR gamma 2, a lipid-activated transcription factor. Cell. 79:1147.
  15. Zhu, Y., Alvares, K., Huang, Q., Rao, M. S. and Reddy, J. K. 1993. Cloning of a new member of the peroxisome proliferator-activated receptor gene family from mouse liver. J Bioi Chem. 268:26817.

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  1. Relationship between Single Nucleotide Polymorphisms in the Peroxisome Proliferator-Activated Receptor Gamma Gene and Fatty Acid Composition in Korean Native Cattle vol.29, pp.2, 2016, https://doi.org/10.5713/ajas.15.0502