Identification and functional characterizations of a novel TRIF gene from grass carp (Ctenopharyngodon idella)

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	Identification and functional characterizations of a novel TRIF gene from grass carp (Ctenopharyngodon idella)
	Yang, Chunrong 1; Li, Qingmei 1; Su, Jianguo 2; Chen, Xiaohui 2; Wang, Yaping 3; Peng, Limin 2
刊名	DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
	2013-10-01
卷号	41 期号:2 页码:222-229
关键词	Grass carp (Ctenopharyngodon idella) TRIF Genomic structure Promoter activity Antiviral immunity Grass carp reovirus
ISSN号	0145-305X
通讯作者	Su, JG (reprint author), Northwest A&F Univ, Coll Anim Sci & Technol, Yangling 712100, Peoples R China.
中文摘要	Toll/interleukin-1 receptor (TIR) domain containing adapter inducing interferon-beta (TRIF) is an adapter in responding to activation of some toll-like receptors (TLRs), which provides early clearance of viral and bacterial pathogens. Here we identified and characterized a full-length genomic sequence of TRIF gene from grass carp Ctenophatyngodon idella (designated as CiTRIF). CiTRIF genomic sequence consists of 3534 base pairs (bp), containing 5' flank sequence (496 bp) and unique intron (815 bp). The full-length cDNA sequence is 2241 bp, including 5' untranslated region (UTR) of 352 bp, 3' UTR of 209 bp, and an open reading frame of 1680 bp encoding a polypeptide of 559 amino acids with an estimated molecular weight of 62.643 kDa and a predicted isoelectric point of 5.71. The deduced amino acid sequence just contains TIR domain, and is most similar to the zebrafish (Danio rerio) TRIF sequence with an identity of 64%. CiTRIF exhibits sequence divergence from its orthologs. Promoter region was predicted and promoter activity was verified. mRNA expression of CiTRIF gene is widespread in 15 tissues investigated, highly in foregut and skin physiological immune barrier. The transcripts of CiTRIF were significantly and rapidly induced in spleen and head kidney tissues at early stage post grass carp reovirus (GCRV) challenge. The modulations are significant but mild in CIK (C idella kidney) cells post GCRV infection or poly(I:C) stimulation. The over-expression vector was constructed and transfected into CIK cell line to get stably expressing recombinant proteins. In CiTRIF transfected cells, mRNA expressions of CiTRIF, CiRIG-I, CiIRF7 and CilFN-I were up-regulated. After GCRV infection, the transcripts of CiTRIF, CiRIG-I, CiIRF7 and CifFN-I fell a little bit after a rapidly and strongly rise. In CiTRIF over-expression cells, virus load and titer were significantly lower than those in controls post GCRV challenge, and virus replication was inhibited obviously. The results indicate that the novel TRIF gene from grass carp plays important roles in modulating antiviral innate immune responses, and serve the further functional studies on TRIF gene in teleosts and immune evolution. (C) 2013 Elsevier Ltd. All rights reserved.
英文摘要	Toll/interleukin-1 receptor (TIR) domain containing adapter inducing interferon-beta (TRIF) is an adapter in responding to activation of some toll-like receptors (TLRs), which provides early clearance of viral and bacterial pathogens. Here we identified and characterized a full-length genomic sequence of TRIF gene from grass carp Ctenophatyngodon idella (designated as CiTRIF). CiTRIF genomic sequence consists of 3534 base pairs (bp), containing 5' flank sequence (496 bp) and unique intron (815 bp). The full-length cDNA sequence is 2241 bp, including 5' untranslated region (UTR) of 352 bp, 3' UTR of 209 bp, and an open reading frame of 1680 bp encoding a polypeptide of 559 amino acids with an estimated molecular weight of 62.643 kDa and a predicted isoelectric point of 5.71. The deduced amino acid sequence just contains TIR domain, and is most similar to the zebrafish (Danio rerio) TRIF sequence with an identity of 64%. CiTRIF exhibits sequence divergence from its orthologs. Promoter region was predicted and promoter activity was verified. mRNA expression of CiTRIF gene is widespread in 15 tissues investigated, highly in foregut and skin physiological immune barrier. The transcripts of CiTRIF were significantly and rapidly induced in spleen and head kidney tissues at early stage post grass carp reovirus (GCRV) challenge. The modulations are significant but mild in CIK (C idella kidney) cells post GCRV infection or poly(I:C) stimulation. The over-expression vector was constructed and transfected into CIK cell line to get stably expressing recombinant proteins. In CiTRIF transfected cells, mRNA expressions of CiTRIF, CiRIG-I, CiIRF7 and CilFN-I were up-regulated. After GCRV infection, the transcripts of CiTRIF, CiRIG-I, CiIRF7 and CifFN-I fell a little bit after a rapidly and strongly rise. In CiTRIF over-expression cells, virus load and titer were significantly lower than those in controls post GCRV challenge, and virus replication was inhibited obviously. The results indicate that the novel TRIF gene from grass carp plays important roles in modulating antiviral innate immune responses, and serve the further functional studies on TRIF gene in teleosts and immune evolution. (C) 2013 Elsevier Ltd. All rights reserved.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Immunology ; Zoology
研究领域[WOS]	Immunology ; Zoology
关键词[WOS]	NF-KAPPA-B ; DOMAIN-CONTAINING ADAPTER ; TOLL-LIKE RECEPTOR-3 ; INTERFERON ANTIVIRAL RESPONSE ; IMMUNE-RESPONSES ; RECOGNITION RECEPTORS ; PATHOGEN RECOGNITION ; INNATE IMMUNITY ; BETA TRIF ; PATHWAY
收录类别	SCI
资助信息	National Key Technology Research and Development Program of China [2012BAD25B0201-04]; Program for New Century Excellent Talents in University [NCET-08-0466]; Chinese Universities Scientific Fund [QN2009022]
语种	英语
WOS记录号	WOS:000323870800013
公开日期	2014-01-06
内容类型	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/342005/19572]
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.Northwest A&F Univ, Coll Vet Med, Yangling 712100, Peoples R China 2.Northwest A&F Univ, Coll Anim Sci & Technol, Yangling 712100, Peoples R China 3.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
推荐引用方式 GB/T 7714	Yang, Chunrong,Li, Qingmei,Su, Jianguo,et al. Identification and functional characterizations of a novel TRIF gene from grass carp (Ctenopharyngodon idella)[J]. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,2013,41(2):222-229.
APA	Yang, Chunrong,Li, Qingmei,Su, Jianguo,Chen, Xiaohui,Wang, Yaping,&Peng, Limin.(2013).Identification and functional characterizations of a novel TRIF gene from grass carp (Ctenopharyngodon idella).DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,41(2),222-229.
MLA	Yang, Chunrong,et al."Identification and functional characterizations of a novel TRIF gene from grass carp (Ctenopharyngodon idella)".DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 41.2(2013):222-229.