Abstract:Abstract Peroxisome proliferator-activated receptor γ (PPARγ) is a ligand-activated nuclear transcriptional factor, which plays an important role in regulating lipid metabolism. This study was carried out to clone the full-length cDNA of pigeon (Columba livia) PPARγ gene, analyze the protein structures, physical, chemical properties and their functional domain structures which encoded by PPARγ gene, and reveal its tissue expression pattern. In our experiment, rapid-amplification cDNA ends (RACE) was used to obtain pigeon PPARγ cDNA. The structure and function of PPARγ gene was analyzed by bioinformatics methods. Real-time PCR was performed to detect PPARγ mRNA expression levels in various tissues. The results showed that the full-length of pigeon PPARγ (GenBank No. KU166859.2) cDNA was of 1 846 bp including a 1 428 bp ORF, 63 bp 5'UTR and 355 bp 3'UTR. The ORF encoded a polypeptide of 475 amino acids, with a calculated relative molecular mass of 54.438 79 kD, pI of 6.19 and an average of hydropathicity -0.287. The predicted protein of pigeon PPARγ was composed of 7 661 atoms with the formula of C2436H3840N644O714S27. Structure prediction showed that there were 2 zinc finger structures and a ligand binding domain in the pigeon PPARγ protein. The deduced amino acid sequence of pigeon PPARγ shared 97%, 97%, 96%, 96%, 96%, 96%, 96%, 92%, 91%, 90% and 64% identities with those of Cuculus canorus, Anas platyrhynchos, Melopsittacus undulates, Taeniopygia guttata, Meleagris gallopavo, Gallus gallus, Coturnix japonica, Sus scrofa, Homo sapien, Mus musculus and Danio rerio, repectively. Phylogenetic relationship analysis indicated that all birds and mammals each formed a large branch, and zebrafish formed another independent branch. Columba livia was closely related with Cuculus canorus and Anas platyrhynchos, which was similar to the results of the biological classification. Real-time PCR analysis revealed that the mRNA level of pigeon PPARγ was highest in lung and abundant in kidney, liver, spleen and heart, while relatively low in muscle. The results showed the full-length cDNA sequence, molecular characteristics and tissue-specific distribution of pigeon PPARγ, which suggested that the PPARγ gene might have various effects on different tissues of pigeion, and also provide theoretical foundation for the further study of the structure and function of PPARγ.
[1]Braissant O, Foufelle F, Scotto C, et al.Differential expression of peroxisome proliferator-activated receptors (PPARs): tissue distribution of PPAR-alpha, -beta, and -gamma in the adult rat[J].Endocrinology, 1996, 137(1):354-366
[2]Bullwinkle EM, Parker MD, Bonan NF, et al.Adipocytes contribute to the growth and progression of multiple myeloma: Unraveling obesity related differences in adipocyte signaling[J].Cancer letters, 2016, 380(1):114-121
[3]Chuang CH, Yeh CL, Yeh SL, et al.Quercetin metabolites inhibit MMP-2 expression in A549 lung cancer cells by PPAR-γ associated mechanisms[J].The Journal of nutritional biochemistry, 2016, 33(1):45-53
[4]Desvergne B, IJpenberg A, Devchand PR, et al.The peroxisome proliferator-activated receptors at the cross-road of diet and hormonal signalling[J].The Journal of steroid biochemistry and molecular biology, 1998, 65(1-6):65-74
[5]Fu RQ, Liu RR, Zhao GP, et al.Expression profiles of key transcription factors involved in lipid metabolism in Beijing-You chickens[J].Gene, 2014, 537(1):120-125
[6]Grindflek E, Sundvold H, Klungland H, et al.Characterisation of porcine peroxisome proliferator-activated receptors gamma 1 and gamma 2: detection of breed and age differences in gene expression[J].Biochemical and biophysical research communications, 1998, 249(3):713-718
[7]Guo F, Ren X, Dong Y, et al.Constitutive expression of PPARγ inhibits proliferation and migration of gastric cancer cells and down-regulates Wnt/β-Catenin signaling pathway downstream target genes TERT and ENAH[J].Gene, 2016, 584(1):31-37
[8]Hummasti S, Tontonoz P.The peroxisome proliferator-activated receptor N-terminal domain controls isotype-selective gene expression and adipogenesis[J].Molecular endocrinology, 2006, 20(6):1261-1275
[9]Ji S, Yang R, Lu C, et al.Differential Expression of PPARγ, FASN, and ACADM Genes in Various Adipose Tissues and Longissimus dorsi Muscle from Yanbian Yellow Cattle and Yan Yellow Cattle[J].Asian-Australasian Journal of Animal Sciences, 2014, 27(1):10-18
[10]Jia Y, Wu C, Kim J, et al.Astaxanthin reduces hepatic lipid accumulations in high-fat-fed C57BL/6J mice via activation of peroxisome proliferator-activated receptor (PPAR) alpha and inhibition of PPAR gamma and Akt[J].The Journal of nutritional biochemistry, 2016, 28(1):9-18
[11]Kawai M, Green CB, Lecka-Czernik B, et al.A circadian-regulated gene, Nocturnin, promotes adipogenesis by stimulating PPAR-gamma nuclear translocation[J].Proceedings of the National Academy of Sciences of the United States of America, 2010, 107(23):10508-10513
[12]Lecarpentier Y, Vallée A.Opposite Interplay between PPAR Gamma and Canonical Wnt/Beta-Catenin Pathway in Amyotrophic Lateral Sclerosis[J].Frontiers in neurology, 2016, 7(Article 100):1-7
[13]Lehrke M, Lazar MA.The many faces of PPARgamma[J].Cell, 2005, 123(6):993-999
[14]Liu HJ, Liao HH, Yang Z, et al.Peroxisome Proliferator-Activated Receptor-γ Is Critical to Cardiac Fibrosis[J].Peroxsimone proliferator-activated receptors research, 2016, 2016(Article ID 2198645):1-12
[15]Liu FC, Tsai YF, Yu HP.Maraviroc attenuates trauma- hemorrhage- induced hepatic injury through PPAR gamma-dependent pathway in rats[J].PLoS one, 2013, 8(10):e78861-e78861
[16]Luo JB,Tian Y,Tao ZR.Regulatory Expression of Peroxisome Proliferator Activated Receptors Genes During Fatty Liver Formation in Geese [J].Agricultural Sciences in China, 2010, 9(1):113-120
[17]Meng H, Li H, Zhao JG, et al.Differential expression of peroxisome proliferator- activated receptors alpha and gamma gene in various chicken tissues[J].Domestic Animal Endocrinology, 2005, 28(1):105-110
[18]Michalik L, Devergne B, Dreyer C, et al.PPAR expression and function during vertebrate development[J].The International journal of developmental biology, 2002, 46(1)(1):105-114
[19]Momose H, Kaminua T, Tanaka Y.Analysis of gene regulation net work by nuelear receptor PPAR[J].Genome Imformatics, 2003, 14(1):364-365
[20]Motoki T, Kurobe H, Hirata Y, et al.PPAR-γ agonist attenuates inflammation in aortic aneurysm patients[J].General thoracic and cardiovascular surgery, 2015, 63(10):565-571
[21]Mueller E.Understanding the variegation of fat; novel regulators of adipocyte differentiation and fat tissue biology[J].Biochimica et biophysica acta, 2014, 1842(3):352-357
[22]Oku H, Umino T.Molecular characterization of peroxisome proliferator-activated receptors (PPARs) and their gene expression in the differentiating adipocytes of red sea bream Pagrus major[J].Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology, 2008, 151(3):268-277
[23]Park S, Kim da S, Kang S.Vitamin D deficiency impairs glucose-stimulated insulin secretion and increases insulin resistance by reducing PPAR-γ expression in nonobese Type 2 diabetic rats[J].The Journal of nutritional biochemistry, 2016, 27(1):257-265
[24]Sakamoto Y, Kanatsu J, Toh M, et al.The Dietary Isoflavone Daidzein Reduces Expression of Pro-Inflammatory Genes through PPARα/γ and JNK Pathways in Adipocyte and Macrophage Co-Cultures[J].PLoS one, 2016, 11(2):e0149676-e0149676
[25]Sato K, Fukao K, Seki Y, et al.Expression of the chicken peroxisome proliferator- activated receptor- gamma gene is influenced by aging, nutrition, and agonist administration[J].Poultry science, 2004, 83(8):1342-1347
[26]Semple RK, Chatterjee VK, O'Rahilly S.PPAR gamma and human metabolic disease[J].The Journal of clinical investigation, 2006, 116(3):581-589
[27]Shchelkunova TA, Morozov IA, Rubtsov PM, et al.Lipid regulators during atherogenesis: expression of LXR, PPAR, and SREBP mRNA in the human aorta[J].PLoS one, 2013, 8(5):e63374-e63374
[28]Taniguchi M, Guan LL, Zhang B, et al.Adipogenesis of bovine perimuscular preadipocytes[J].Biochemical and biophysical research communications, 2008, 366(1):54-59
[29]Tontonoz P, Spiegelman BM.Fat and beyond: the diverse biology of PPARgamma[J].Annual review of biochemistry, 2008, 77(1):289-312
[30]Velleman SG, Coy CS, Emmerson DA.Effect of the timing of posthatch feed restrictions on the deposition of fat during broiler breast muscle development[J].Poultry science, 2014, 10(93):2622-2627
[31]Walczak R, Tontonoz P.PPARadigms and PPARadoxes: expanding roles for PPARgamma in the control of lipid metabolism[J].Journal of lipid research, 2002, 43(2):177-186
[32]Wang L, Na W, Wang YX et al.Characterization of chicken PPARγ expression and its impact on adipocyte proliferation and differentiation[J].Yi Chuan, 2012, 34(4):454-464
[33]Wu Y, Liu X, Xiao H, et al.The differential expression of peroxisome proliferators-activated receptors in various duck tissues[J].Molecular biology reports, 2010, 37(3):(3):1235-1240
[34]Zhou Y, Wu Y, Qin Y, et al.Ampelopsin Improves Insulin Resistance by Activating PPARγ and Subsequently Up-Regulating FGF21-AMPK Signaling Pathway[J].PLoS one, 2016, 11(7):e0159191-e0159191