A tailless Dscam from Eriocheir sinensis diversified by alternative splicing

doi:10.1016/j.fsi.2013.04.029

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	A tailless Dscam from Eriocheir sinensis diversified by alternative splicing
	Wang, Jingjing1,2 ; Wang, Lingling 1; Gao, Yang1,2 ; Jiang, Qiufen 1,2; Yi, Qilin 1,2; Zhang, Huan 1; Zhou, Zhi 1; Qiu, Limei 1; Song, Linsheng 1; Song, LS
	2013-08-01
发表期刊	FISH & SHELLFISH IMMUNOLOGY
ISSN	1050-4648
卷号	35 期号:2 页码:249-261
文章类型	Article
摘要	Dscam (Down syndrome cell adhesion molecule), a member of the immunoglobulin superfamily (IgSF), plays an essential role in pathogen recognition and further involves in the innate defense of invertebrates. In the present study, the cDNA of a Dscam from Chinese mitten crab Eriocheir sinensis (designated EsDscam) was cloned and characterized. It contained a 5-terminal untranslated region (UTR) of 60 bp, a 3-UTR of 216 bp with a poly (A) tail, and an open reading frame (ORF) of 4848 bp encoding a polypeptide of 1615 amino acids with the putative molecular mass of 178.4 kDa and theoretical isoelectric point of 6.31. The EsDscam protein shared higher sequence identities and similar domain architecture with Dscams from other invertebrate, including typical 10 immunoglobulin (Ig) domains, 6 fibronectin type 3 domains (FNIII) and one cell attachment sequence (RGD) in extracellular region, while it lacked the expected transmembrane domain and cytoplasmic tail compared with other members of Dscam family. After sequencing 80 separate clones of Ig2, 3 and Ig7 regions from pooled cDNA libraries constructed from normal and bacterial-infected crabs, 44 alternative sequences were detected in the N-terminal of Ig2, 39 ones in Ig3, and 31 ones in Ig7 domain, suggesting that EsDscam could potentially encode at least 53196 unique isoforms. Furthermore, two 3'UTR isoforms and two 5'UTR isoforms of EsDscam were also identified by RACE strategy. EsDscam mRNA was most abundantly expressed in the tissues of nerve, muscle, hepatopancreas and gill, and weakly expressed in heart, gonad and hemocytes. Western blotting and immunofluorescence analysis revealed that EsDscam protein was mainly distributed in serum, and few on the membrane of crab hemocytes. These results suggested that this tailless EsDscam was one member of crustacean Dscam family, and the generation of diverse isoforms through alternative splicing allowed it to recognize various pathogens and play an active role in immune defense of crabs. (C) 2013 Elsevier Ltd. All rights reserved.; Dscam (Down syndrome cell adhesion molecule), a member of the immunoglobulin superfamily (IgSF), plays an essential role in pathogen recognition and further involves in the innate defense of invertebrates. In the present study, the cDNA of a Dscam from Chinese mitten crab Eriocheir sinensis (designated EsDscam) was cloned and characterized. It contained a 5-terminal untranslated region (UTR) of 60 bp, a 3-UTR of 216 bp with a poly (A) tail, and an open reading frame (ORF) of 4848 bp encoding a polypeptide of 1615 amino acids with the putative molecular mass of 178.4 kDa and theoretical isoelectric point of 6.31. The EsDscam protein shared higher sequence identities and similar domain architecture with Dscams from other invertebrate, including typical 10 immunoglobulin (Ig) domains, 6 fibronectin type 3 domains (FNIII) and one cell attachment sequence (RGD) in extracellular region, while it lacked the expected transmembrane domain and cytoplasmic tail compared with other members of Dscam family. After sequencing 80 separate clones of Ig2, 3 and Ig7 regions from pooled cDNA libraries constructed from normal and bacterial-infected crabs, 44 alternative sequences were detected in the N-terminal of Ig2, 39 ones in Ig3, and 31 ones in Ig7 domain, suggesting that EsDscam could potentially encode at least 53196 unique isoforms. Furthermore, two 3'UTR isoforms and two 5'UTR isoforms of EsDscam were also identified by RACE strategy. EsDscam mRNA was most abundantly expressed in the tissues of nerve, muscle, hepatopancreas and gill, and weakly expressed in heart, gonad and hemocytes. Western blotting and immunofluorescence analysis revealed that EsDscam protein was mainly distributed in serum, and few on the membrane of crab hemocytes. These results suggested that this tailless EsDscam was one member of crustacean Dscam family, and the generation of diverse isoforms through alternative splicing allowed it to recognize various pathogens and play an active role in immune defense of crabs. (C) 2013 Elsevier Ltd. All rights reserved.
关键词	Dscam Eriocheir Sinensis Alternative Splicing Immune Defense
学科领域	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
DOI	10.1016/j.fsi.2013.04.029
URL	查看原文
收录类别	SCI
语种	英语
WOS研究方向	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS类目	Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS记录号	WOS:000322207500007
WOS关键词	DROSOPHILA-DSCAM ; NERVOUS-SYSTEM ; IMMUNE-SYSTEM ; CELL-ADHESION ; RECOGNITION ; EVOLUTION ; RECEPTOR ; INSECTS ; VIRUS ; GENES
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
引用统计	被引频次：24[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/16520
专题	实验海洋生物学重点实验室海洋环境腐蚀与生物污损重点实验室
通讯作者	Song, LS
作者单位	1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Wang, Jingjing,Wang, Lingling,Gao, Yang,et al. A tailless Dscam from Eriocheir sinensis diversified by alternative splicing[J]. FISH & SHELLFISH IMMUNOLOGY,2013,35(2):249-261.
APA	Wang, Jingjing.,Wang, Lingling.,Gao, Yang.,Jiang, Qiufen.,Yi, Qilin.,...&Song, LS.(2013).A tailless Dscam from Eriocheir sinensis diversified by alternative splicing.FISH & SHELLFISH IMMUNOLOGY,35(2),249-261.
MLA	Wang, Jingjing,et al."A tailless Dscam from Eriocheir sinensis diversified by alternative splicing".FISH & SHELLFISH IMMUNOLOGY 35.2(2013):249-261.

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