Promoter Amplification, Sequence Comparison of PPARs in Duck (Anas platyrhynchos), and Cluster Analysis with Transcription Factors in Expression Profiles
Abstract：Peroxisome proliferator-activated receptors (PPARs) belong to the nuclear receptor superfamily, and there are 3 subtypes including α, β/δ and γ, and all PPARs members play a crucial role in cell growth and differentiation. In order to investigate their distinct functions in the process of skeletal muscle development, PPARs promoter sequences in duck (Anas platyrhynchos) were cloned using PCR and were analyzed using several bioinformatics tools. The mRNA expression profiles of PPARs and their common transcription factors in the breast and leg muscle of duck during embryonic stage were detected using qRT-PCR. The PCR results showed that PPARα, PPARβ and PPARγ promoter sequences of duck were 2 526, 1 631 and 2 942 bp,respectively, and GenBank accession were KX845431, KX845432 and KX845433, respectively. Their homology reached 99%, 97% and 97% with the predicted sequences of duck genome in NCBI, respectively. All PPARs promoters in duck shared higher homology compared with chicken (Gallus gallus) near the translation initiation sites through analyzing the cloned promoter sequences. The typical cis acting elements (CAAT-box and TATA-box) had been predicted to exist in the promoter region of PPARs, and common binding sites of transcription factors (specificity protein 1 (Sp1), nuclear factor kappa B (NF-κB) and CCAAT/enhancer binding protein alpha (C/EBP-α)) were predicted to exist in all duck PPARs members. Moreover, PPARα and PPARγ shared 9 common transcription factors, and PPARβ shared 8 with PPARγ, while PPARα and PPARβ shared the least number of common transcription factor. The qRT-PCR results showed that PPARα, PPARβ and PPARγ expressed in breast and leg muscle tissues of duck. Clustering results showed that the expression patterns of PPARβ and PPARγ were more similar compared with PPARα in leg muscle. It could be seen that the functions of the 2 genes were more similar in the process of duck skeletal muscle development. Besides, in breast muscle, the expression pattern of transcription factor NF-κB and PPARβ gene was consistent. PPARs members all participated in the regulation of skeletal muscle development, and the function of PPARβ might be larger; the function of PPARβ and PPARγ in skeletal muscle development might be similar. In addition, the NF-κB might control the expression of PPARβ in skeletal muscle. The results provide theoretical basis for the expression and function identification of PPARs gene.
[1]傅小雅, 黄娴.过氧化物酶体增殖剂激活受体的研究进展和应用前景[J].中国热带医学, 2008, 8(2):321-325
[2]李丛丛, 吴艳红, 安利国, 等.PPARδ启动子克隆及活性分析[J].生物化学与生物物理进展, 2007, 34(4):412-417
[3]罗锦标, 袁青妍, 陶争荣, 等.填饲前后鹅 PPARs 基因组织表达特性的研究[J].中国家禽, 2008, 30(15):24-27
[4]马瑶, 程海东.PPAR 基因多态性的研究进展[J].中国优生与遗传杂志, 2007, 15(9):6-9
[5]祁胜宾, 双剑博, 卢晓昭, 等.人GDDR基因启动子的克隆和报告基因载体构建[J].细胞与分子免疫学杂志, 2011, 27(2):229-230
[6]徐洪亮, 赵磊, 卓德祥, 等.NF-κB 对 PPARδ 基因的转录调控[J].中国生物化学与分子生物学报, 2008, 24(10):931-937
[7]Angione A R, Jiang C, Pan D, et al..PPARδ regulates satellite cell proliferation and skeletal muscle regeneration[J].Skeletal muscle, 2011, 1(1):-
[8]Auboeuf D, Rieusset J, Fajas L, et al.Tissue distribution and quantification of the expression of mRNAs of peroxisome proliferator–activated receptors and liver X receptor-α in humans: no alteration in adipose tissue of obese and NIDDM patients[J].Diabetes, 1997, 46(8):1319-1327
[9]Burri L, Thoresen G H, Berge R K.The role of PPAR activation in liver and muscle[J].PPAR research, 2010, :-
[10]Delerive P, De Bosscher K, Besnard S, et al.Peroxisome proliferator-activated receptor α negatively regulates the vascular inflammatory gene response by negative cross-talk with transcription factors NF-κB and AP-1 [J].Journal of Biological Chemistry, 1999, 274(45):32048-32054
[11]Kim S E, Park J H, Cho Y W, et al.Porous chitosan scaffold containing microspheres loaded with transforming growth factor-β1: implications for cartilage tissue engineering[J].Journal of Controlled Release, 2003, 91(3):365-374
[12]Müller F, Demény M A, Tora L.New problems in RNA polymerase II transcription initiation: matching the diversity of core promoters with a variety of promoter recognition factors[J].Journal of Biological Chemistry, 2007, 282(20):14685-14689
[13]Muoio D M, Way J M, Tanner C J, et al.Peroxisome proliferator-activated receptor-α regulates fatty acid utilization in primary human skeletal muscle cells [J].Diabetes, 2002, 51(4):901-909
[14]Wang Y X, Zhang C L, Ruth T Y, et al.Regulation of muscle fiber type and running endurance by PPARδ [J].PLoS Biol, 2004, 2(10):-
[15]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):1235-1240
[16]Yuan G, Chen X, Li D.Modulation of peroxisome proliferator-activated receptor gamma (PPAR γ) by conjugated fatty acid in obesity and inflammatory bowel disease[J].Journal of agricultural and food chemistry, 2015, 63(7):1883-1895
[17]Zhou T, Yan X, Wang G, et al.Evolutionary pattern and regulation analysis to support why diversity functions existed within PPAR gene family members[J].BioMed research international, 2015, :-
