Molecular cloning and characterization of two types of I kappa B alpha orthologues in orange-spotted grouper, Epinephelus coioides
Gao, R; Huang, YH; Huang, XH; Guan, LY; Wei, SN; Zhou, YC; Qin, QW; zychnu@163.com; qinqw@scsio.ac.cn
2014
发表期刊FISH & SHELLFISH IMMUNOLOGY
ISSN1050-4648
卷号38期号:1页码:101-110
摘要Inhibitors of kappa B (I kappa Bs) are the members of primary regulators of NF-kappa B, which can inhibit NF-kappa B activity by blocking the NF-kappa B in an inactive state in the cytoplasm. In this study, two types of IKB alpha orthologues (EcI kappa B alpha A and EcI kappa B alpha B) from orange-spotted grouper, Epinephelus coioides, were cloned and characterized. EcI kappa B alpha A and EcIKB alpha B encoded putative proteins containing 308 and 318 amino acids, which shared 59% and 53% identity to I kappa B alpha A and I kappa B alpha B of Danio rerio, respectively. Amino acid sequence alignment showed that both EcI kappa B alpha A and EcI kappa B alpha B contained a conserved degradation motif DSGLDS in the N-terminal region and a PEST sequence in the C-terminal region. In addition, EcI kappa B alpha A and EcI kappa B alpha B contained 5 and 6 ankcyrin repeats, respectively. The genomic DNA of EcI kappa B alpha A and EcI kappa B alpha B consisted of 6 exons and 5 introns. Both of their transcripts were widely distributed in different tissues, and the expression levels were different in response to various stimuli, including lipopolysaccharide (LPS), Vibrio alginolyticus and Singapore grouper iridovirus (SGIV). Dual-luciferase reporter assay suggested that both EcI kappa B alpha A and EcI kappa B alpha B were able to inhibit Ecc-Rel and Ecp65 induced NF-kappa B promoter activity in grouper spleen (GS) cells. Subcellular localization analysis showed that EcI kappa B alpha B was present predominantly in the cytoplasm, while EclicB alpha A was distributed throughout both the nucleus and the cytoplasm. Furthermore, overexpression of EcI kappa B alpha A and EcI kappa B alpha B in GS cells inhibited the viral gene transcriptions of MCP, ORF019 and ORF162 of SGIV. Taken together, our findings suggested that both EcI kappa B alpha A and EcI kappa B alpha B were involved in grouper innate immunity against virus. (C) 2014 Elsevier Ltd. All rights reserved.
部门归属[Gao, Ren ; Zhou, Yongcan ; Qin, Qiwei] Hainan Univ, Coll Marine Sci, State Key Lab Breeding Base Sustainable Exploitat, Haikou 570228, Peoples R China ; [Gao, Ren ; Huang, Youhua ; Huang, Xiaohong ; Guan, Liya ; Wei, Shina ; Qin, Qiwei] Chinese Acad Sci, South China Sea Inst Oceanol, Key Lab Trop Marine Bioresources & Ecol, Guangzhou 510301, Guangdong, Peoples R China
关键词Epinephelus Coioides Eci Kappa b Alpha a Eci Kappa b Alpha b Nf-kappa b Sgiv
学科领域Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
资助者National Basic Research Program of China (973) [2012CB114402, 2012CB114404]; Natural Science Foundation of China [31172437, 31372566, 31330082]; Knowledge Innovation Program of the Chinese Academy of Sciences [KZCX2-EW-Q213] ; National Basic Research Program of China (973) [2012CB114402, 2012CB114404]; Natural Science Foundation of China [31172437, 31372566, 31330082]; Knowledge Innovation Program of the Chinese Academy of Sciences [KZCX2-EW-Q213] ; National Basic Research Program of China (973) [2012CB114402, 2012CB114404]; Natural Science Foundation of China [31172437, 31372566, 31330082]; Knowledge Innovation Program of the Chinese Academy of Sciences [KZCX2-EW-Q213] ; National Basic Research Program of China (973) [2012CB114402, 2012CB114404]; Natural Science Foundation of China [31172437, 31372566, 31330082]; Knowledge Innovation Program of the Chinese Academy of Sciences [KZCX2-EW-Q213]
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收录类别sci
资助项目LMB
资助者National Basic Research Program of China (973) [2012CB114402, 2012CB114404]; Natural Science Foundation of China [31172437, 31372566, 31330082]; Knowledge Innovation Program of the Chinese Academy of Sciences [KZCX2-EW-Q213] ; National Basic Research Program of China (973) [2012CB114402, 2012CB114404]; Natural Science Foundation of China [31172437, 31372566, 31330082]; Knowledge Innovation Program of the Chinese Academy of Sciences [KZCX2-EW-Q213] ; National Basic Research Program of China (973) [2012CB114402, 2012CB114404]; Natural Science Foundation of China [31172437, 31372566, 31330082]; Knowledge Innovation Program of the Chinese Academy of Sciences [KZCX2-EW-Q213] ; National Basic Research Program of China (973) [2012CB114402, 2012CB114404]; Natural Science Foundation of China [31172437, 31372566, 31330082]; Knowledge Innovation Program of the Chinese Academy of Sciences [KZCX2-EW-Q213]
WOS记录号WOS:000336353700014
引用统计
文献类型期刊论文
条目标识符http://ir.scsio.ac.cn/handle/344004/10444
专题中科院海洋生物资源可持续利用重点实验室
通讯作者zychnu@163.com; qinqw@scsio.ac.cn
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Gao, R,Huang, YH,Huang, XH,et al. Molecular cloning and characterization of two types of I kappa B alpha orthologues in orange-spotted grouper, Epinephelus coioides[J]. FISH & SHELLFISH IMMUNOLOGY,2014,38(1):101-110.
APA Gao, R.,Huang, YH.,Huang, XH.,Guan, LY.,Wei, SN.,...&qinqw@scsio.ac.cn.(2014).Molecular cloning and characterization of two types of I kappa B alpha orthologues in orange-spotted grouper, Epinephelus coioides.FISH & SHELLFISH IMMUNOLOGY,38(1),101-110.
MLA Gao, R,et al."Molecular cloning and characterization of two types of I kappa B alpha orthologues in orange-spotted grouper, Epinephelus coioides".FISH & SHELLFISH IMMUNOLOGY 38.1(2014):101-110.
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