Identification, characterization and the interaction of Tollip and IRAK-1 in grass carp (Ctenopharyngodon idellus)

CORC > 水生生物研究所 > 鱼类生物学及渔业生物技术研究中心 > 期刊论文

	Identification, characterization and the interaction of Tollip and IRAK-1 in grass carp (Ctenopharyngodon idellus)
	Huang, Rong 1; Lv, Jianjian 1,2; Luo, Daji 1; Liao, Lanjie 1; Zhu, Zuoyan 1; Wang, Yaping 1
刊名	FISH & SHELLFISH IMMUNOLOGY
	2012-09-01
卷号	33 期号:3 页码:459-467
关键词	Tollip IRAK-1 Molecular cloning Viral infection Gras g carp
ISSN号	1050-4648
通讯作者	Wang, YP (reprint author), Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, 7 Donghu S Rd, Wuhan 430072, Peoples R China.
中文摘要	Tollip and IRAK-1 are key components of the TLR/IL-1R signaling pathway in mammals, which play crucial roles as mediators of the TLR/IL-1R signal transduction pathways. Although several TLRs have been found in fish, molecular associations, protein protein interactions or the role of the TLR signaling pathway in infection-induced immunity in fish has received little attention. In this study. Tollip and IRAK-1 sequences of grass carp were isolated from a head kidney cDNA library. Full length transcripts and sequences of promoter regions were obtained by 3' and 5' RACE and genome walking, respectively. Reporter gene-promoter constructs and real-time RT-PCR analysis was used to determine grass carp Tollip and IRAK-1 transcription pattern in tissues. Recombinant proteins were used for antibodies production. Phylogenetically, the grass carp loci clustered with previously reported Tollip and IRAK-1genes, respectively, and their sequences shared the highest identity with the genes of zebrafish (Danio rerio). The promoter region of grass carp Tollip and IRAK-1 proved to be active. After viral infection transcript levels of both loci were upregulated in most immune-related tissues in a time-dependent manner. Using antibodies produced in this study, immunofluorescence analysis indicated that Tollip and IRAK-1 were uniformly distributed and co-localized in the cytoplasm of CIK cells. After viral infection, however, Tollip and IRAK-1 both trended toward the cell membrane. Our results demonstrate the existence of Tollip and IRAK-1 proteins in teleost species, and suggest that Tollip-IRAK-1 complexes are being recruited to receptor complexes after stimulation with virus. These results provide novel insights into the role of the TLR signaling pathway in teleosts, especially the action of teleost Tollip and IRAK-1 and the interaction of these molecules as part of this pathway. (C) 2012 Elsevier Ltd. All rights reserved.
英文摘要	Tollip and IRAK-1 are key components of the TLR/IL-1R signaling pathway in mammals, which play crucial roles as mediators of the TLR/IL-1R signal transduction pathways. Although several TLRs have been found in fish, molecular associations, protein protein interactions or the role of the TLR signaling pathway in infection-induced immunity in fish has received little attention. In this study. Tollip and IRAK-1 sequences of grass carp were isolated from a head kidney cDNA library. Full length transcripts and sequences of promoter regions were obtained by 3' and 5' RACE and genome walking, respectively. Reporter gene-promoter constructs and real-time RT-PCR analysis was used to determine grass carp Tollip and IRAK-1 transcription pattern in tissues. Recombinant proteins were used for antibodies production. Phylogenetically, the grass carp loci clustered with previously reported Tollip and IRAK-1genes, respectively, and their sequences shared the highest identity with the genes of zebrafish (Danio rerio). The promoter region of grass carp Tollip and IRAK-1 proved to be active. After viral infection transcript levels of both loci were upregulated in most immune-related tissues in a time-dependent manner. Using antibodies produced in this study, immunofluorescence analysis indicated that Tollip and IRAK-1 were uniformly distributed and co-localized in the cytoplasm of CIK cells. After viral infection, however, Tollip and IRAK-1 both trended toward the cell membrane. Our results demonstrate the existence of Tollip and IRAK-1 proteins in teleost species, and suggest that Tollip-IRAK-1 complexes are being recruited to receptor complexes after stimulation with virus. These results provide novel insights into the role of the TLR signaling pathway in teleosts, especially the action of teleost Tollip and IRAK-1 and the interaction of these molecules as part of this pathway. (C) 2012 Elsevier Ltd. All rights reserved.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
研究领域[WOS]	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
关键词[WOS]	RECEPTOR-ASSOCIATED KINASE ; INTERLEUKIN-1 RECEPTOR ; IL-1 RECEPTOR ; PARALICHTHYS-OLIVACEUS ; SIGNALING PATHWAYS ; NEGATIVE REGULATOR ; MOLECULAR-CLONING ; JAPANESE FLOUNDER ; GENE-EXPRESSION ; INNATE IMMUNITY
收录类别	SCI
资助信息	National Natural Science Foundation of China [31130055]; National Basic Research Program of China [2009CB118701]
语种	英语
WOS记录号	WOS:000308057700001
公开日期	2013-01-09
内容类型	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/342005/17228]
专题	水生生物研究所_鱼类生物学及渔业生物技术研究中心_期刊论文
作者单位	1.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
推荐引用方式 GB/T 7714	Huang, Rong,Lv, Jianjian,Luo, Daji,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.
APA	Huang, Rong,Lv, Jianjian,Luo, Daji,Liao, Lanjie,Zhu, Zuoyan,&Wang, Yaping.(2012).Identification, characterization and the interaction of Tollip and IRAK-1 in grass carp (Ctenopharyngodon idellus).FISH & SHELLFISH IMMUNOLOGY,33(3),459-467.
MLA	Huang, Rong,et al."Identification, characterization and the interaction of Tollip and IRAK-1 in grass carp (Ctenopharyngodon idellus)".FISH & SHELLFISH IMMUNOLOGY 33.3(2012):459-467.