绵羊UCP1基因碱基突变与其蛋白结构和功能

doi:10.3864/j.issn.0578-1752.2012.14.021

摘要/Abstract

摘要： 【目的】检测两个绵羊群体中解偶联蛋白1（uncoupling protein 1，UCP1）基因的单核苷酸多态性（single nucleotide polymorphism，SNP），预测和分析碱基改变对蛋白结构和功能的影响。【方法】利用PCR-SSCP结合测序的方法检测UCP1基因编码区的SNPs，用生物信息学方法分析UCP1蛋白的理化性质和结构。【结果】UCP1基因编码区存在4个SNPs，其中c.214G＞A（Val72Met）和c.273C＞T位于外显子2，c.624C＞T和c.757G＞A（Ala253Thr）位于外显子5。进一步分析导致氨基酸改变的c.214G＞A和c.757 G＞A突变发现，此两个突变只对UCP1蛋白的理化性质和转录因子结合位点有细微改变，未引起UCP1蛋白空间构象的变化，对蛋白表达的影响不大。【结论】基于对人类中两个与肥胖相关的外显子突变型蛋白结构的比较说明，UCP1蛋白结构的改变可能影响其功能。

关键词: 绵羊, 解偶联蛋白1（UCP1）, 单核苷酸多态性（SNP）, 生物信息学, 蛋白结构

Abstract: 【Objective】The objectives of this study are to detect the single nucleotide polymorphism (SNP) of UCP1 gene in two fat-tailed sheep breeds, and to predict the protein structure and function caused by the mutations. 【Method】 PCR-SSCP combined with sequencing were used to detect the SNPs in coding region of UCP1 gene, and bioinformatics approach was used to predict the physicochemical properties and structures of UCP1 protein. 【Result】 Four SNPs existed in the coding region, with c.214G＞A (Val72Met) and c.273C＞T located on exon 2, c.624C＞T and and c.757G＞A (Ala253Thr) on exon 5. Further analysis indicated that the two mutations which caused amino acid substitution had little influence on the protein expression. This was because of the two mutations merely resulted in a slight change of the physicochemical properties and transcription factor binding site, but did not lead to the change of its spatial configuration. 【Conclusion】Comparison of UCP1 protein structures between two exonic mutants, which associated respectively with human obesity and diabetes, and the normal structure suggested that the changes in UCP1 protein structure could influence the protein function.

Key words: sheep, uncoupling protein 1 (UCP1), single nucleotide polymorphism (SNP), bioinformatics, protein structure

袁亚男, 刘文忠, 刘建华, 乔利英, 武建亮. 绵羊UCP1基因碱基突变与其蛋白结构和功能[J]. 中国农业科学, 2012, 45(14): 2973-2980.

YUAN Ya-Nan, LIU Wen-Zhong, LIU Jian-Hua, QIAO Li-Ying, WU Jian-Liang. The Base Mutation of Ovine UCP1 Gene and Its Relationship with Protein Structure and Function[J]. Scientia Agricultura Sinica, 2012, 45(14): 2973-2980.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2012.14.021

https://www.chinaagrisci.com/CN/Y2012/V45/I14/2973

参考文献

[1]Clarke L, Bryant M J, Lomax M A, Symonds M E. Maternal manipulation of brown adipose tissue and liver development in the ovine fetus during late gestation. The British Journal of Nutrition. 1997, 77(6): 871-883.

[2]Bassett J M, Bomford J, Mott J C. Photoperiod: an important regulator of plasma prolactin concentration in fetal lambs during late gestation. Quarterly Journal of Experimental Physiology, 1988, 73(2): 241-244.

[3]Sale M M, Hsu F C, Palmer N D, Gordon C J, Keene K L, Borgerink H M, Sharma A J, Bergman R N, Taylor K D, Saad M F, Norris J M. The uncoupling protein 1 gene, UCP1, is expressed in mammalian islet cells and associated with acute insulin response to glucose in African American families from the IRAS family study. BMC Endocrine Disorders, 2007, 7: 1-10.

[4]Rudofsky G Jr, Schrodter A, Voron’ko O E, Schlotterer A, Humpert P M, Tafel J, Nawroth P P, Bierhaus A, Hamann A. Promoter polymorphisms of UCP1, UCP2, and UCP3 are not associated with diabetic microvascular complications in type 2 diabetes. Hormone and Metablic Research, 2007, 39(4): 306-309.

[5]Kim K S, Cho D Y, Kim Y J, Choi S M, Kim J Y, Shin S U, Yoon Y S. The finding of new genetic polymorphism of UCP-1 A-1766G and its effects on body fat accumulation. Biochimica et Biophysica Acta, 2005, 1741(1/2): 149-155.

[6]Shin H D, Kim K S, Cha M H, Yoon Y. The effects of UCP-1 polymorphisms on obesity phenotypes among Korean female subjects. Biochemical and Biophysical Research Communications, 2005, 335(2): 624-630.

[7]Nagai N, Sakane N, Kotani K, Hamada T, Tsuzaki K, Moritani T. Uncoupling protein 1 gene -3826 A/G polymorphism is associated with weight loss on a short-term, controlled-energy diet in young women. Nutrition Research, 2011, 31: 255-261.

[8]Mori H, Okazawa H, Iwamoto K, Maeda E, Hashiramoto M, Kasuga M. A polymorphism in the 5′ untranslated region and a Met229--＞Leu variant in exon 5 of the human UCP1 gene are associated with susceptibility to type II diabetes mellitus. Diabetologia, 2001, 44(3): 373-376.

[9]Labruna G, Pasanisi F, Fortunato G, Nardelli C, Finelli C, Farinaro E, Contaldo F, Sacchetti L. Sequence analysis of the UCP1 gene in a severe obese population from Southern Italy. Journal of Obesity, 2011. doi: 10.1155/2011/269043.

[10]Hu J, Zhou H, Smyth A, Luo Y, Hickford J G. Polymorphism of the bovine ADRB3 gene. Molecular Biology Reports, 2010, 37(7): 3389-3392.

[11]Walker J M. The Proteomics Protocols Handbook. Totowa, N.J: Humana Press, 2005: 988.

[12]Cheng J, Saigo H, Baldi P. Large-scale prediction of disulphide bridges using kernel methods, two-dimensional recursive neural networks, and weighted graph matching. Proteins, 2006, 62(3): 617-629.

[13]Randall A, Cheng J, Sweredoski M, Baldi P. TMBpro: secondary structure, beta-contact and tertiary structure prediction of transmembrane beta-barrel proteins. Bioinformatics, 2008, 24(4): 513-520.

[14]Rost B, Yachdav G, Liu J. The PredictProtein server. Nucleic Acids Research, 2004, 32: 321-326.

[15]Petersen T N, Brunak S, von Heijne G, Nielsen H. SignalP 4.0: discriminating signal peptides from transmembrane regions. Nature Methods, 2011, 8: 785-786.

[16]Julenius K, Molgaard A, Gupta R, Brunak S. Prediction, conservation analysis, and structural characterization of mammalian mucin-type O-glycosylation sites. Glycobiology, 2005, 15(2): 153-164.

[17]Blom N, Gammeltoft S, Brunak S. Sequence and structure-based prediction of eukaryotic protein phosphorylation sites. Journal of Molecular Biology, 1999, 294(5): 1351-1362.

[18]Heinemeyer T, Wingender E, Reuter I, Hermjakob H, Kel A E, Kel O V, Ignatieva E V, Ananko E A, Podkolodnaya O A, Kolpakov F A, Podkolodny N L, Kolchanov N A. Databases on transcriptional regulation: TRANSFAC, TRRD and COMPEL. Nucleic Acids Research, 1998, 26(1): 362-367.

[19]Arnold K, Bordoli L, Kopp J, Schwede T. The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics, 2006, 22(2): 195-201.

[20]Kiefer F, Arnold K, Kunzli M, Bordoli L, Schwede T. The SWISS- MODEL repository and associated resources. Nucleic Acids Research, 2009, 37: D387-D392.

[21]Kopp J, Schwede T. The SWISS-MODEL repository of annotated three-dimensional protein structure homology models. Nucleic Acids Research, 2004, 32: D230-D234.

[22]Benkert P, Biasini M, Schwede T. Toward the estimation of the absolute quality of individual protein structure models. Bioinformatics, 2011, 27: 343-350.

[23]Tamura K, Dudley J, Nei M, Kumar S. MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 2007, 24(8): 1596-1599.

[24]Kozak L P, Britton J H, Kozak U C, Wells J M. The mitochondrial uncoupling protein gene. Correlation of exon structure to transmembrane domains. Journal of Biological Chemistry, 1988, 263: 12274-12277.

[25]Yubero P, Manchado C, Cassard-Doulcier A M, Mampel T, Vinas O, Iglesias R, Giralt M, Villarroya F. CCAAT/enhancer binding proteins alpha and beta are transcriptional activators of the brown fat uncoupling protein gene promoter. Biochemical and Biophysical Research Communications, 1994, 198(2): 653-659.

[26]Kumar N, Liu D, Wang H, Robidoux J, Collins S. Orphan nuclear receptor NOR-1 enhances 3′, 5′-cyclic adenosine 5′-monophosphate- dependent uncoupling protein-1 gene transcription. Molecular Endocrinology, 2008, 22(5): 1057-1064.

[27]Wang H, Zhang Y, Yehuda-Shnaidman E, Medvedev A V, Kumar N, Daniel K W, Robidoux J, Czech M P, Mangelsdorf D J, Collins S. Liver X receptor alpha is a transcriptional repressor of the uncoupling protein 1 gene and the brown fat phenotype. Molecular and Cellular Biology, 2008, 28(7): 2187-2200.

[28]Shore A, Karamitri A, Kemp P, Speakman J R, Lomax M A. Role of Ucp1 enhancer methylation and chromatin remodelling in the control of Ucp1 expression in murine adipose tissue. Diabetologia, 2010, 53(6): 1164-1173.

[29]Yuan Y N, Liu W Z, Liu J H, Qiao L Y, Wu J L. Cloning and ontogenetic expression of the uncoupling protein 1 gene (UCP1) in sheep. Journal of Applied Genetics, 2012, 53(2): 203-212.