坝上长尾鸡MITF基因核心启动子鉴定与分析
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摘要
旨在探明坝上长尾鸡小眼畸形相关转录因子(microphthalmia-associated transcription factor,MITF)基因启动子活性区与结构,为研究MITF基因在毛囊组织中的表达调控提供依据。本研究采用双荧光素酶表达载体的方法,构建了6个含有不同长度坝上长尾鸡MITF基因启动子片段的pGL3-Basic表达载体及4个核心启动子区突变载体,转染DF1细胞48h后检测双荧光素酶活性。结果,确定了鸡MITF基因启动子的核心区域为-660~+200bp。其中突变位点-579bp、-505bp、-274bp、-220bp、-203~-202bp、-98bp、-46bp、-14bp、+1bp、+28bp对MITF基因启动子活性有较大影响,在突变区域预测到HOX家族(HOXA3、HOXD8、HOXD9、HOXD10、HOX11)、NF-1、DBP、TFIID和TFIIB与MITF序列的结合性发生了变化。
The research was designed to study the activity and the structure of MITFpromoter and to provide clues for studying this gene's regulation mechanism in hair follicle.Dual-luciferase expression vectors were constructed and transfected to DF1 cells with lip2000.The dual-luciferase detection kit was used to measure the relative luciferase activity.The 6 expression vectors with different promoter regions of MITFgene and 4 mutant vectors of the core promoter region were constructed in Bashang Long-tail chicken.The core promoter region from-660 bp to+200 bp was identified in chicken MITFgene.The function of-579 bp,-505 bp,-274 bp,-220 bp,-203--202 bp,-98 bp,-46 bp,-14 bp,+1 bp and+28 bp sites in regulating promoter activity was critical.The similarities of transcription factor binding sites including HOX family(HOXA3,HOXD8,HOXD9,HOXD10 and HOX11),NF-1,DBP,TFIID and TFIIB to MITF were changed predicted by software.
The research was designed to study the activity and the structure of MITFpromoter and to provide clues for studying this gene's regulation mechanism in hair follicle.Dual-luciferase expression vectors were constructed and transfected to DF1 cells with lip2000.The dual-luciferase detection kit was used to measure the relative luciferase activity.The 6 expression vectors with different promoter regions of MITFgene and 4 mutant vectors of the core promoter region were constructed in Bashang Long-tail chicken.The core promoter region from-660 bp to+200 bp was identified in chicken MITFgene.The function of-579 bp,-505 bp,-274 bp,-220 bp,-203--202 bp,-98 bp,-46 bp,-14 bp,+1 bp and+28 bp sites in regulating promoter activity was critical.The similarities of transcription factor binding sites including HOX family(HOXA3,HOXD8,HOXD9,HOXD10 and HOX11),NF-1,DBP,TFIID and TFIIB to MITF were changed predicted by software.
引文
[1]赵兵令,王海东,陈天直,等.小鼠毛囊不同生长周期中MITF下游色素相关基因的定位表达及相关性分析[J].中国生物化学与分子生物学报,2017,33(2):198-206.ZHAO B L,WANG H D,CHEN T Z,et al.Localization and correlation analysis of MITF downstream pigmentation-related genes at different growth cycle of mouse hair follicle[J].Chinese Journal of Biochemistry and Molecular Biology,2017,33(2):198-206.(in Chinese)
[2]VACHTENHEIM J,BOROVANSKY J."Transcription physiology"of pigment formation in melanocytes:central role of MITF[J].Exp Dermatol,2010,19(7):617-627.
[3]STEINGRIMSSON E,COPELAND N G,JENKINS N A.Melanocytes and the Microphthalmia transcription factor network[J].Annu Rev Genet,2004,38(38):365-411.
[4]黎钊,王平,洪为松,等.正常人黑素细胞MITF对酪氨酸酶相关蛋白的转录调控研究[J].医学研究杂志,2013,42(3):58-62.LI Z,WANG P,HONG W S,et al.Effect of MITF transcriptional control on family of tyrosinase-related proteins in normal human melanocytes[J].Journal of Medical Research,2013,42(3):58-62.(in Chinese)
[5]杨玉静,张丹瑾,聂瑞强,等.绵羊MITF-M在黑素细胞中过表达后的功能分析[J].中国农业科学,2016,49(21):4214-4221.YANG Y J,ZHANG D J,NIE R Q,et al.The function analysis of over-expression of oar MITF-M in melanocytes[J].Scientia Agricultura Sinica,2016,49(21):4214-4221.(in Chinese)
[6]郑嫩珠,辛清武,朱志明,等.白绒乌鸡MITF基因的cDNA克隆、表达及其对黑色素沉积的影响[J].中国农业科学,2015,48(18):3711-3718.ZHENG N Z,XIN Q W,ZHU Z M,et al.cDNA cloning and expression of MITF gene and its effect on melanin deposition in silky fowl[J].Scientia Agricultura Sinica,2015,48(18):3711-3718.(in Chinese)
[7]MARKAKIS M N,SOEDRING V E,DANTZER V,et al.Association of MITFgene with hearing and pigmentation phenotype in Hedlund white American mink(Neovison vison)[J].J Genet,2014,93(2):477-481.
[8]牛晓艳,任克良,曹亮,等.獭兔MITF基因部分外显子多态性研究[J].山西农业科学,2013,41(11):1231-1234.NIU X Y,REN K L,CAO L,et al.Research of polymorphisms of parts of exons in MITF gene in rex rabbit[J].Journal of Shanxi Agricultural Sciences,2013,41(11):1231-1234.(in Chinese)
[9]HARTMAN M L,CZYZ M.MITF in melanoma:mechanisms behind its expression and activity[J].Cell Mol Life Sci,2015,72(7):1249-1260.
[10]张廷焕.猪MITF基因转录调控区域克隆及活性分析[D].雅安:四川农业大学,2014.ZHANG T H.Cloning and activity analysis of pig microphthalmia-associtated transcription factor(MITF)gene promoter[D].Ya'an:Sichuan Agricultural University,2014.(in Chinese)
[11]KNUTSON B A,HAHN S.TFIIB-related factors in RNA polymerase I transcription[J].Biochim Biophys Acta,2013,1829(3-4):265-273.
[12]LIU X,BUSHNELL D A,WANG D,et al.Structure of an RNA polymerase II-TFIIB complex and the transcription initiation mechanism[J].Science,2010,327(5962):206-209.
[13]ORPHANIDES G,LAGRANGE T,REINBERG D.The general transcription factors of RNA polymerase II[J].Genes Dev,1996,10(21):2657-2683.
[14]ROEDER R G.The role of general initiation factors in transcription by RNA polymerase II[J].Trends Biochem Sci,1996,21(9):327-335.
[15]张爱贤,夏云飞.普通转录因子TFⅡB与肝细胞癌相关的研究进展[J].南通大学学报:医学版,2011,31(1):58-61.ZHANG A X,XIA Y F.Advances in research on the association of general transcription factor II B and hepatocellular carcinoma[J].Journal of Nantong University:Medical Sciences,2011,31(1):58-61.(in Chinese)
[16]KATSUMA S.Transcription of dbp from the coding region of the Bm17gene is required for the efficient propagation of Bombyx mori nucleopolyhedrovirus[J].Virus Res,2016,223:57-63.
[17]吕杨,陈蕾,谢院生,等.转录因子DBP对系膜增殖性肾炎免疫炎症反应的作用及其机制研究[C]∥中国中西医结合学会肾脏疾病专业委员会2015年学术年会资料汇编.南宁:中国中西医结合学会肾脏疾病专业委员会,2015:846.LV Y,CHEN L,XIE Y S,et al.The role of transcriptional factor DBP in immune inflammatory response to mesangial proliferative nephritis and its mechanism[C]∥Information Compilation of the 2015Academic Annual Meeting of the Professional Committee on Kidney Disease in Chinese and Western Medicine Society.Nanning:Chinese Academy of Integrated Traditional Chinese and Western Medicine Kidney Disease Specialized Committee,2015:846.(in Chinese)
[18]BABAJKO S,GROYER A.Interplay of the liver-enriched trans-acting factors,DBP and HNF1,in the transactivation of human IGFBP-1promoter[J].Biochem Biophys Res Commun,1993,196(1):480-486.
[19]张涛,张子平,贾锡伟,等.杂色鲍HSP90基因启动子的功能分析[J].水产学报,2017,41(4):490-497.ZHANG T,ZHANG Z P,JIA X W,et al.Functional analysis of the promoter in Haliotis diversicolor HSP90gene[J].Journal of Fisheries of China,2017,41(4):490-497.(in Chinese)
[20]李丽莎,彭永东,郑晓宁,等.山羊PMEL基因启动子活性及转录调控元件分析[J].畜牧兽医学报,2017,48(5):826-835.LI L S,PENG Y D,ZHENG X N,et al.Analysis of the promoter activity and transcriptional regulatory elements of goat PMELgene[J].Acta Veterinaria et Zootechnica Sinica,2017,48(5):826-835.(in Chinese)
[21]MORGAN R,SIMPSON G,GRAY S,et al.HOX transcription factors are potential targets and markers in malignant mesothelioma[J].BMC Cancer,2016,16(1):85-96.
[22]金淑清,浦予飞,裘莹.HOX基因的研究进展[J].癌症进展,2011,9(2):154-158.JIN S Q,PU Y F,QIU Y.Hox genes mechanisms[J].Oncology Progress,2011,9(2):154-158.(in Chinese)
[23]李慧,花保祯.Hox基因及其进化机制的研究进展[J].动物学杂志,2011,46(1):136-142.LI H,HUA B Z.Hox genes and their evolutionary mechanisms[J].Chinese Journal of Zoology,2011,46(1):136-142.(in Chinese)
[2]VACHTENHEIM J,BOROVANSKY J."Transcription physiology"of pigment formation in melanocytes:central role of MITF[J].Exp Dermatol,2010,19(7):617-627.
[3]STEINGRIMSSON E,COPELAND N G,JENKINS N A.Melanocytes and the Microphthalmia transcription factor network[J].Annu Rev Genet,2004,38(38):365-411.
[4]黎钊,王平,洪为松,等.正常人黑素细胞MITF对酪氨酸酶相关蛋白的转录调控研究[J].医学研究杂志,2013,42(3):58-62.LI Z,WANG P,HONG W S,et al.Effect of MITF transcriptional control on family of tyrosinase-related proteins in normal human melanocytes[J].Journal of Medical Research,2013,42(3):58-62.(in Chinese)
[5]杨玉静,张丹瑾,聂瑞强,等.绵羊MITF-M在黑素细胞中过表达后的功能分析[J].中国农业科学,2016,49(21):4214-4221.YANG Y J,ZHANG D J,NIE R Q,et al.The function analysis of over-expression of oar MITF-M in melanocytes[J].Scientia Agricultura Sinica,2016,49(21):4214-4221.(in Chinese)
[6]郑嫩珠,辛清武,朱志明,等.白绒乌鸡MITF基因的cDNA克隆、表达及其对黑色素沉积的影响[J].中国农业科学,2015,48(18):3711-3718.ZHENG N Z,XIN Q W,ZHU Z M,et al.cDNA cloning and expression of MITF gene and its effect on melanin deposition in silky fowl[J].Scientia Agricultura Sinica,2015,48(18):3711-3718.(in Chinese)
[7]MARKAKIS M N,SOEDRING V E,DANTZER V,et al.Association of MITFgene with hearing and pigmentation phenotype in Hedlund white American mink(Neovison vison)[J].J Genet,2014,93(2):477-481.
[8]牛晓艳,任克良,曹亮,等.獭兔MITF基因部分外显子多态性研究[J].山西农业科学,2013,41(11):1231-1234.NIU X Y,REN K L,CAO L,et al.Research of polymorphisms of parts of exons in MITF gene in rex rabbit[J].Journal of Shanxi Agricultural Sciences,2013,41(11):1231-1234.(in Chinese)
[9]HARTMAN M L,CZYZ M.MITF in melanoma:mechanisms behind its expression and activity[J].Cell Mol Life Sci,2015,72(7):1249-1260.
[10]张廷焕.猪MITF基因转录调控区域克隆及活性分析[D].雅安:四川农业大学,2014.ZHANG T H.Cloning and activity analysis of pig microphthalmia-associtated transcription factor(MITF)gene promoter[D].Ya'an:Sichuan Agricultural University,2014.(in Chinese)
[11]KNUTSON B A,HAHN S.TFIIB-related factors in RNA polymerase I transcription[J].Biochim Biophys Acta,2013,1829(3-4):265-273.
[12]LIU X,BUSHNELL D A,WANG D,et al.Structure of an RNA polymerase II-TFIIB complex and the transcription initiation mechanism[J].Science,2010,327(5962):206-209.
[13]ORPHANIDES G,LAGRANGE T,REINBERG D.The general transcription factors of RNA polymerase II[J].Genes Dev,1996,10(21):2657-2683.
[14]ROEDER R G.The role of general initiation factors in transcription by RNA polymerase II[J].Trends Biochem Sci,1996,21(9):327-335.
[15]张爱贤,夏云飞.普通转录因子TFⅡB与肝细胞癌相关的研究进展[J].南通大学学报:医学版,2011,31(1):58-61.ZHANG A X,XIA Y F.Advances in research on the association of general transcription factor II B and hepatocellular carcinoma[J].Journal of Nantong University:Medical Sciences,2011,31(1):58-61.(in Chinese)
[16]KATSUMA S.Transcription of dbp from the coding region of the Bm17gene is required for the efficient propagation of Bombyx mori nucleopolyhedrovirus[J].Virus Res,2016,223:57-63.
[17]吕杨,陈蕾,谢院生,等.转录因子DBP对系膜增殖性肾炎免疫炎症反应的作用及其机制研究[C]∥中国中西医结合学会肾脏疾病专业委员会2015年学术年会资料汇编.南宁:中国中西医结合学会肾脏疾病专业委员会,2015:846.LV Y,CHEN L,XIE Y S,et al.The role of transcriptional factor DBP in immune inflammatory response to mesangial proliferative nephritis and its mechanism[C]∥Information Compilation of the 2015Academic Annual Meeting of the Professional Committee on Kidney Disease in Chinese and Western Medicine Society.Nanning:Chinese Academy of Integrated Traditional Chinese and Western Medicine Kidney Disease Specialized Committee,2015:846.(in Chinese)
[18]BABAJKO S,GROYER A.Interplay of the liver-enriched trans-acting factors,DBP and HNF1,in the transactivation of human IGFBP-1promoter[J].Biochem Biophys Res Commun,1993,196(1):480-486.
[19]张涛,张子平,贾锡伟,等.杂色鲍HSP90基因启动子的功能分析[J].水产学报,2017,41(4):490-497.ZHANG T,ZHANG Z P,JIA X W,et al.Functional analysis of the promoter in Haliotis diversicolor HSP90gene[J].Journal of Fisheries of China,2017,41(4):490-497.(in Chinese)
[20]李丽莎,彭永东,郑晓宁,等.山羊PMEL基因启动子活性及转录调控元件分析[J].畜牧兽医学报,2017,48(5):826-835.LI L S,PENG Y D,ZHENG X N,et al.Analysis of the promoter activity and transcriptional regulatory elements of goat PMELgene[J].Acta Veterinaria et Zootechnica Sinica,2017,48(5):826-835.(in Chinese)
[21]MORGAN R,SIMPSON G,GRAY S,et al.HOX transcription factors are potential targets and markers in malignant mesothelioma[J].BMC Cancer,2016,16(1):85-96.
[22]金淑清,浦予飞,裘莹.HOX基因的研究进展[J].癌症进展,2011,9(2):154-158.JIN S Q,PU Y F,QIU Y.Hox genes mechanisms[J].Oncology Progress,2011,9(2):154-158.(in Chinese)
[23]李慧,花保祯.Hox基因及其进化机制的研究进展[J].动物学杂志,2011,46(1):136-142.LI H,HUA B Z.Hox genes and their evolutionary mechanisms[J].Chinese Journal of Zoology,2011,46(1):136-142.(in Chinese)