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钙激活氯离子通道调节剂(calcium-activated chloride channel regulator, CLCA)为一类金属蛋白酶依赖家族,在哺乳动物上皮组织杯状细胞的粘液产生和粘液平衡中起重要作用。为了探究CLCA1基因在凡纳滨对虾渗透压中的作用机制,本研究采用RACE技术在凡纳滨对虾中克隆出了CLCA1基因,该基因总长度为3 129 bp,5'端非编码区长度为175 bp,3'端非编码区长度为107 bp,开放阅读框ORF长度为2 847 bp,共编码984个氨基酸,有1个跨膜区域,膜外有VMA结构域。与其他物种氨基酸序列比对结果显示,凡纳滨对虾CLCA1氨基酸与其他物种相似性都较低,相似度最高的为刀额新对虾(Procambarus clarkii) CLCA2氨基酸序列,为47.83%。RT-qPCR结果显示,CLCA1基因在凡纳滨对虾各组织中均有表达,其中肠道、肝胰腺和鳃中的表达量较高;对不同盐度下凡纳滨对虾4个组织中CLCA1基因的定量结果显示,随着盐度的下降,CLCA1基因的表达量在4个组织中呈下调趋势,表明CLCA1基因与凡纳滨对虾的渗透压调节相关。本研究初步阐明了CLCA1基因的理化性质,对CLCA1基因在凡纳滨对虾体内各组织和不同盐度下各组织的表达定量可为CLCA1基因在今后甲壳动物的渗透压调节及免疫调节的研究提供基础支持。
Abstract：Calcium-activated chloride channel regulator(CLCA) is a metalloproteinase-dependent family protein and plays an essential role in goblet cell mucus production and balance from the epithelium in mammals. In order to figure out the osmotic pressure regulation of CLCA1 in Litopenaeus vannamei, we cloned full-length of CLCA1 cDNA by RACE technology. The total length of the gene was 3 129 bp, with the length of the 5'-terminal non-coding region of 175 bp, the length of the 3'-terminal non-coding region of 107 bp, and the ORF length of the open reading frame of 2 847 bp. It encoded 984 amino acids in total, with one transmembrane region, and a VMA structure domain outside the membrane. The results of amino acid sequence comparison with other species showed that the CLCA1 amino acid of Litopenaeus vannamei was not similar to the other species, the highest similarity was the CLCA2 amino acid sequence of Procambarus clarkii, which was 47.83%. RT-qPCR results showed that CLCA1 gene was expressed in all tissues of Litopenaeus vannamei, the expressions were higher in intestine, gill and hepatopancreas. Results showed the expression of CLCA1 gene was down-regulated in four tissues as salinity decreased which meant that CLCA1 was involved in osmotic pressure regulation of Litopenaeus vannamei. The experiment initially clarified the physical and chemical properties of CLCA1 gene and quantified CLCA1 gene expression in different tissues of litopenaeus vannamei under different salinity. These results provided basic support for future studies on the role of CLCA1 gene in osmotic pressure regulation in crustaceans.
Ale vy Y.G.,Patel A.C.,Romero A.G.,Patel D.A.,Tucker J.,Roswit W.T.,Miller C.A.,Heier R.F.,Byers D.E.,Brett T.J.,and Holtzman M.J.,2012,IL-13-induced airway mucus production is attenuated by MAPK13 inhibition,J.Clin.Invest.,122(12):4555-4568.
Becker A.K.,Mikolajek H.,Paulsson M.,Wagener R.,and Werner J.M.,2014,A structure of a collagenⅥVWA domain displays N and C termini at opposite sides of the protein,Structure,22(2):199-208.
Bothe M.K.,Mundhenk L.,Kaup M.,Weise C.,and Gruber A.D.,2011,The murine goblet cell protein m CLCA3 is a zinc-dependent metalloprotease with autoproteolytic activity,Mol.Cells,32(6):535-541.
Ching J.C.H.,Lobanova L.,and Loewen M.E.,2013,Secreted h CLCA1 is a signaling molecule that activates airway macrophages,PLo S ONE,8(12):e83130.
Cunningham S.A.,Awayda M.S.,Bubien J.K.,Ismailov I.I.,Arrate M.P.,Berdiev B.K.,Benos D.J.,and Fuller C.M.,1995,Cloning of an epithelial chloride channel from bovine trachea,J.Biol.Chem.,270(52):31016-31026.
Dolphin A.C.,2016,Voltage-gated calcium channels and their auxiliary subunits:physiology and pathophysiology and pharmacology,J.Physiol.,594(19):5369-5390.
Erickson N.A.,Dietert K.,Enders J.,Glauben R.,Nouailles G.,Gruber A.D.,and Mundhenk L.,2018,Soluble mucus component CLCA1 modulates expression of leukotactic cytokines and BPIFA1 in murine alveolar macrophages,but not in bone marrow-derived macrophages,Histochem.Cell Biol.,149(6):619-633.
Lacy D.B.,Wigelsworth D.J.,Melnyk R.A.,Harrison S.C.,and Collier R.J.,2004a,Structure of heptameric protective antigen bound to an anthrax toxin receptor:a role for receptor in p H-dependent pore formation,Proc.Natl.Acad.Sci.US-A,101(36):13147-13151.
Lacy D.B.,Wigelsworth D.J.,Scobie H.M.,Young J.A.,and Collier R.J.,2004b,Crystal structure of the von willebrand factor A domain of human capillary morphogenesis protein 2:an anthrax toxin receptor,Proc.Natl.Acad.Sci.USA,101(17):6367-6372.
Mall M.A.,Danahay H.,and Boucher R.C.,2018,Emerging concepts and therapies for mucoobstructive lung disease,Ann.Am.Thorac.Soc.,15(S3):216-226.
Mundhenk L.,Johannesson B.,Anagnostopoulou P.,Braun J.,Bothe M.K.,Schultz C.,Mall M.A.,and Gruber A.D.,2012,MCLCA3 does not contribute to calcium-activated chloride conductance in murine airways,Am.J.Respir.Cell Mol.Biol.,47(1):87-93.
Nystr觟m E.E.L.,Arike L.,Ehrencrona E.,Hansson G.C.,and Johansson M.E.V.,2019,Calcium-activated chloride channel regulator 1 (CLCA1) forms non-covalent oligomers in colonic mucus and has mucin 2-processing properties,Cell Biol.,294(45):17075-17089.
Patel A.C.,Brett T.J.,and Holtzman M.J.,2009,The role of CL-CA proteins in inflflammatory airway disease,Annu.Rev.Physiol.,71:425-449.
Ponce-Palafox J.T.,Martinez-Palacios C.A.,and Ross L.G.,1997,The effects of salinity and temperature on the growth and survival rates of juvenile white shrimp,Penaeus vannamei,Boone,1931,Aquaculture,157(1):107-115.
Poole A.,Urbanek C.,Eng C.,Schageman J.,Jacobson S.,Brian P.O.,Galanter J.M.,Gignoux C.R.,Roth L.A.,Kumar J.,Lutz S.,Liu A.H.,Fingerlin T.E.,Setterquist R.A.,Burchard E.G.,Santana J.R.,and Seibold M.A.,2014,Dissecting childhood asthma with nasal transcriptomics distinguishes subphenotypes of disease,J.Allergy Clin.Immunol.,133(3):670-678.
Ruffifin M.,Voland M.,Marie S.,Bonora M.,Blanchard E.,Blouquit L.S.,Naline E.,Puyo P.,Le R.P.,Guillot L.,Harriet C.H.,Clement A.,and Tabary O.,2013,Anoctamin 1dysregulation alters bronchial epithelial repair in cystic fibrosis,Biochim.Biophys.Acta,1832(12):2340-2351.
Sala R.M.,Yurtsever Z.,Berry K.N.,Nichols C.G.,and Brett T.J.,2017,Modulation of TMEM16A channel activity by the von Willebrand factor type A (VWA) domain of the calcium activated chloride channel regulator 1 (CLCA1),J.Biol.Chem.,292(22):9164-9174.
Santelli E.,Bankston L.A.,Leppla S.H.,and Liddington R.C.,2004,Crystal structure of a complex between anthrax toxin and its host cell receptor,Nature,430(7002):905-908.
Saoud I.P.,Davis D.A.,and Rouse D.B.,2003,Suitability studies of inland well waters for Litopenaeus vannamei culture,Aquaculture,217(1-4):373-383.
Shimaoka M.,Takagi J.,and Springer T.A.,2002,Conformational regulation of integrin structure and function,Annu.Rev.Biophys.Biomol.Struct.,31:485-516.
Song G.J.,Koksal A.C.,Lu C.F.,and Springer T.A.,2012,Shape change in the receptor for gliding motility in Plasmodium sporozoites,Proc.Natl.Acad.Sci.USA,109(52):21420-21425.
Song G.J.,and Springer T.A.,2014,Structures of the Toxoplasma gliding motility adhesin,Proc.Natl.Acad.Sci.USA,111(13):4862-4867.
Sonneville F.,Ruffifin M.,Coraux C.,Rousselet N.,Le Rouzic P.,Blouquit Laye S.,Corvol H.,and Tabary O.,2017,Micro R-NA-9 downregulates the ANO1 chloride channel and contributes to cystic fibrosis lung pathology,Nat.Commun.,8(1):710-721.
Springer T.A.,2006,Complement and the multifaceted functions of VWA and integrin I domains,Structure,14(11):1611-1616.
Veit G.,Bossard F.,Goepp J.,Verkman A.S.,Galietta L.J.,Hanrahan J.W.,and Lukacs G.L.,2012,Proinflflammatory cytokine secretion is suppressed by TMEM16A or CFTRchannel activity in human cystic fibrosis bronchial epithelia,Mol.Biol.Cell,23(21):4188-4202.
Wang Q.,and Liu F.Q.,2000,Studies on the pathogens and pathogenesis of black gill diseases of Penaeus chinensis,Dongwu Kexue yu Dongwu Yixue (Animal Science&Veterinary Medicine),17(5):48-50.(王茜,刘凤岐,2000,中国对虾黑鳃病病原及病变机理的研究,动物科学与动物医学,17(5):48-50.)
Yu H.,Li Q.,Kolosov V.P.,Perelman J.M.,and Zhou X.,2010,Interleukin-13 induces mucin 5AC production involving STAT6/SPDEF in human airway epithelial cells,Cell Commun.Adhes.,17(4-6):83-92.
Yurtsever Z.,Rabanal M.S.,Randolph D.T.,Scheaffer S.M.,Roswit W.T.,Alevy Y.G.,Patel A.C.,Heier R.F.,Romero A.G.,Nichols C.G.,Holtzman M.J.,and Brett T.J.,2012,Self-cleavage of human CLCA1 protein by a novel internal metalloprotease domain controls calcium-activated chloride channel activation,J.Biol.Chem.,287(50):42138-42149.
Zhou Y.H.,Dong Q.,Louahed J.,Dragwa C.,Savio D.,Huang M.X.,Weiss C.,Tomer Y.,Mc Lane M.P.,Nicolaides N.C.,and Levitt R.C.,2001,Characterization of a calcium-activated chloride channel as a shared target of Th2 cytokine pathways and its potential involvement in asthma,Am.J.Respir.Cell Mol.Biol.,25(4):486-491.
基本信息:
DOI：10.13417/j.gab.040.001067
中图分类号:S917.4
引用信息:
[1]温贝妮,刘红,赵宇,等.凡纳滨对虾(Litopenaeus vannamei)盐度调控相关基因CLCA1的克隆及表达分析[J].基因组学与应用生物学,2021,40(03):1067-1076.DOI:10.13417/j.gab.040.001067.
基金信息:
上海市科技兴农项目(2019-02-08-00-10-F01111)资助
2021-03-25
2021-03-25