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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
Source PublicationFISH & SHELLFISH IMMUNOLOGY
ISSN1050-4648
Volume38Issue:1Pages:101-110
AbstractInhibitors 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.
Department[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
KeywordEpinephelus Coioides Eci Kappa b Alpha a Eci Kappa b Alpha b Nf-kappa b Sgiv
Subject AreaFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
Funding OrganizationNational 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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Indexed Bysci
Funding ProjectLMB
Funding OrganizationNational 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 IDWOS:000336353700014
Citation statistics
Cited Times:12[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.scsio.ac.cn/handle/344004/10444
Collection中科院海洋生物资源可持续利用重点实验室
Corresponding Authorzychnu@163.com; qinqw@scsio.ac.cn
Recommended Citation
GB/T 7714
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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