黄郁葱,蔡双虎,蔡佳,简纪常,吴灶和,蔡小辉

• 1:广东海洋大学水产学院
• 2:广东省水产经济动物病原生物学及流行病学重点实验室
• 3:仲恺农业工程学院
• 4:广西海洋生物技术重点实验室

摘要(Abstract)：

用同源克隆和RACE的方法克隆红笛鲷(Lutjanus sanguineus)白细胞介素1受体相关激酶1(Interleukin-1receptor-associated kinase 1,IRAK-1)基因的cDNA全序列,并用荧光定量PCR分析其在健康鱼及哈维弧菌感染后红笛鲷的组织表达。结果表明,该序列全长3 207 bp(登录号KF728204),包含5′非编码区(UTR)202 bp,3′UTR752 bp,开放阅读框(ORF)2 253 bp,编码750个氨基酸。根据推导的氨基酸序列预测其蛋白分子质量为82.6 ku,理论等电点为6.59。IRAK-1包含1个N端死亡结构域、proST结构域、中央激酶结构域和C末端结构域。荧光定量PCR分析显示,红笛鲷IRKA-1基因在肝脏和皮肤的表达量最高,其次是心脏、鳃、肌肉和胸腺,其余组织的表达量较低。哈维弧菌侵染红笛鲷后,各组织中IRAK-1基因mRNA表达量均呈上调趋势,肝组织中变化最显著。

关键词(KeyWords)： 红笛鲷;IRAK-1;基因克隆;组织表达

Abstract:

Keywords:

基金项目(Foundation): 广东省自然基金(2016A030313748);; 广西海洋生物技术重点实验室开放基金(GLMBT-201404);; 广东省海洋经济创新发展区域示范专项(GD2012-B01-004);; 广东省科技计划(2012A020800006);; 大学生创新创业训练计划(CXXL2014020)

作者(Author): 黄郁葱,蔡双虎,蔡佳,简纪常,吴灶和,蔡小辉

参考文献(References)：

• [1]Janeway CA Jr,Medzhitov R.Innate immune recognition[J].Annual Review of Immunology,2002,20(1):197-216.
• [2]KAWAI T,AKIRA S.The role of pattern-recognition receptors in innate immunity:update on Toll-like receptors[J].Nature Immunology,2010,11(5):373-384.
• [3]AKIRA S,TAKEDA K,KAISHO T.Toll-like receptors:critical proteins linking innate and acquired immunity[J].Nature Immunology,2001,2(8):675-680.
• [4]SCHNARE M,BARTON G M,HOLT A C,et al.Toll-like receptors control activation of adaptive immune responses[J].Nature Immunology,2001,2(10):947-950.
• [5]TAKEDA K,AKIRA S.Toll-like receptors[J].Current Protocols in Immunology,2015,109(Suppl.):14.12.1-14.12.10.doi:10.1002/0471142735.im1412s109.
• [6]JANSSENS S,BEYAERT R.Functional diversity and regulation of different interleukin-1 receptor-associated kinase(IRAK)family members[J].Molecular Cell,2003,11(2):293-302.
• [7]FLANNERY S,BOWIE A G.The interleukin-1receptor-associated kinases:critical regulators of innate immune signalling[J].Biochemical Pharmacology,2010,80(12):1981-1991.
• [8]CAO Z D,HENZEL WJ,GAO X.IRAK:a kinase associated with the interleukin-1 receptor[J].Science,1996,271(5252):1128-1131.
• [9]TAKEDA K,AKIRA S.TLR signaling pathways[J].Seminars in Immunology,2004,16(1):3-9.
• [10]GAN L,LI L W.Regulations and roles of the interleukin-1 receptor associated kinases(IRAKs)in innate and adaptive immunity[J].Immunologic Research,2006,35(3):295-302.
• [11]KANAKARAJ P,NGO K,WU Y,et al.Defective interleukin(IL)-18-mediated natural killer and T helper cell type 1 responses in IL-1 receptor-associated kinase(IRAK)-deficient mice[J].The Journal of Experimental Medicine,1999,189(7):1129-1138.
• [12]THOMAS J A,ALLEN J L,TSEN M,et al.Impaired cytokine signaling in mice lacking the IL-1receptor-associated kinase[J].The Journal of Immunology,1999,163(2):978-984.
• [13]NGUYEN H,CHATTERJEE-KISHORE M,JIANG Z F,et al.IRAK-dependent phosphorylation of Stat1 on serine 727 in response to interleukin-1 and effects on gene expression[J].Journal of Interferon&Cytokine Research,2003,23(4):183-192.
• [14]UEMATSU S,SATO S,YAMAMOTO M,et al.Interleukin-1 receptor-associated kinase-1 plays an essential role for Toll like receptor(TLR)7-and TLR9-mediated interferon-αinduction[J].The Journal of Experimental Medicine,2005,201(6):915-923.
• [15]HUANG Y S,LI T,SANE D C,et al.IRAK1 serves as a novel regulator essential for lipopolysaccharide-induced interleukin-10 gene expression[J].The Journal of Biological Chemistry,2004,279(49):51697-51703.
• [16]ZHANG C Z,YIN Z X,HE W,et al.Cloning of IRAK1and its upregulation in symptomatic mandarin fish infected with ISKNV[J].Biochemical and Biophysical Research Communications,2009,383(3):298-302.
• [17]HUANG R,LV J J,LUO D J,et al.Identification,characterization and the interaction of Tollip and IRAK-1 in grass carp(Ctenopharyngodon idellus)[J].Fish&Shellfish Immunology,2012,33(3):459-467.
• [18]LI Y W,ZHAO F,MO Z Q,et al.Characterization,expression,and functional study of IRAK-1 from grouper,Epinephelus coioides[J].Fish&Shellfish Immunology,2016,56:374-381.
• [19]XU Y H,HUANG Y C,CAI S H.Characterization and function analysis of interleukin-1 receptor-associated kinase-1(IRAK-1)from Fenneropenaeus penicillatus[J].Fish&Shellfish Immunology,2017,61:111-119.
• [20]ZHANG X Z,DAI L P,WU Z H,et al.Expression pattern of heat shock protein 90 gene of humphead snapper Lutjanus sanguineus during pathogenic Vibrio harveyi stress[J].Journal of Fish Biology,2011,79(1):178-193.
• [21]PHELAN P E,MELLON M T,KIM C H.Functional characterization of full-length TLR3,IRAK-4,and TRAF6 in zebrafish(Danio rerio)[J].Molecular Immunology,2005,42(9):1057-1071.
• [22]黄郁葱,鲁义善,简纪常,等.红笛鲷IRAK-4基因c DNA全长的克隆及组织表达分析[J].广东海洋大学学报,2015,35(1):18-27.
• [23]Li Y W,Mo X B,Zhou L,et al.Identification of IRAK-4in grouper(Epinephelus coioides)that impairs My D88-dependent NF-κB activation[J].Developmental&Comparative Immunology,2014,45(1):190-197.
• [24]NEUMANN D,KOLLEWE C,RESCH K,et al.The death domain of IRAK-1:an oligomerization domain mediating interactions with My D88,Tollip,IRAK-1,and IRAK-4[J].Biochemical and Biophysical Research Communications,2007,354(4):1089-1094.
• [25]LIN S C,LO Y C,WU H.Helical assembly in the My D88-IRAK4-IRAK2 complex in TLR/IL-1Rsignalling[J].Nature,2010,465(7300):885-890.
• [26]MARTIN M U,KOLLEWE C.Interleukin-1receptor-associated kinase-1(IRAK-1):A self-regulatory adapter molecule in the signaling cascade of the Toll/IL-1 receptor family[J].Signal Transduction,2015,1(1/2):37-50.
• [27]YE H,ARRON J R,LAMOTHE B,et al.Distinct molecular mechanism for initiating TRAF6 signalling[J].Nature,2002,418(6896):443-447.
• [28]LI X X,COMMANE M,JIANG Z F,et al.IL-1-induced NFκB and c-Jun K activation diverge at IL-1receptor-associated kinase(IRAK)[J].Proceedings of the National Academy of Sciences of the United States of America,2001,98(8):4461-4465.
• [29]KOLLEWE C,MACKENSEN A C,NEUMANN D,et al.Sequential autophosphorylation steps in the interleukin-1 receptor-associated kinase-1 regulate its availability as an adapter in interleukin-1 signaling[J].The Journal of Biological Chemistry,2004,279(7):5227-5236.
• [30]TROFIMOVA M,SPRENKLE A B,GREEN M,et al.Developmental and tissue-specific expression of mouse pelle-like protein kinase[J].The Journal of Biological Chemistry,1996,271(30):17609-17612.
• [31]BERCZI I,SZENTIVANYI A.The acute phase response[J].Neuroimmune Biology,2003,3:463-494.
• [32]SANCéAU J,KAISHO T,HIRANO T,et al.Triggering of the human interleukin-6 gene by interferon-γand tumor necrosis factor-αin monocytic cells involves cooperation between interferon regulatory factor-1,NFκB,and Sp1 transcription factors[J].The Journal of Biological Chemistry,1995,270(46):27920-27931
• [33]MUKAIDA N,MAHE Y,MATSUSHIMA K.Cooperative interaction of nuclear factor-kappa B,and cis-regulatory enhancer binding protein-like factor binding elements in activating the interleukin-8 gene by pro-inflammatory cytokines[J].The Journal of Biological Chemistry,1990,265(34):21128-21133.
• [34]Uribe C,FOLCH H U,ENRIQUEZ R U,et al.Innate and adaptive immunity in teleost fish:a review[J].VeterinárníMedicína,2011,56(10):486-503.
• [35]LAZADO CC,CAIPANG CM.Mucosal immunity and probiotics in fish[J].Fish and Shellfish Immunology,2014,39(1):78-89.