Molecular cloning and functional characterization of peptidoglycan recognition protein 6 in grass carp Ctenopharyngodon idella

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

	Molecular cloning and functional characterization of peptidoglycan recognition protein 6 in grass carp Ctenopharyngodon idella
	Li, Jun Hua 1,2; Yu, Zhang Long 1,2; Xue, Na Na 1,2; Zou, Peng Fei 1,2; Hu, Jing Yu 1; Nie, P.1; Chang, Ming Xian 1
刊名	DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
	2014
卷号	42 期号:2 页码:244-255
关键词	Peptidoglycan recognition protein PGRP6 Peptidoglycan-binding activity Antibacterial activity Grass carp Ctenopharyngodon idella
ISSN号	0145-305X
通讯作者	Chang, MX (reprint author), Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei Province, Peoples R China.
中文摘要	Peptidoglycan recognition proteins (PGRPs) are pattern recognition molecules of innate immunity. In this study, a long-form PGRP, designated as gcPGRP6, was identified from grass carp Ctenopharyngodon idella. The deduced amino acid sequence of gcPGRP6 is composed of 464 residues with a conserved PGRP domain at the C-terminus. The gcPGRP6 gene consists of four exons and three introns, spacing approximately 2.7 kb of genomic sequence. Phylogenetic analysis demonstrated that gcPGRP6 is clustered closely with zebrafish PGLYRP6, and formed a long-type PGRP subfamily together with PGLYRP2 members identified in teleosts and mammals. Real-time PCR and Western blotting analyses revealed that gcPGRP6 is constitutively expressed in organs/tissues examined, and its expression was significantly induced in liver and intestine of grass carp in response to PGN stimulation and in CIK cells treated with lipoteichoic acid (LTA), polyinosinic polycytidylic acid (Poly I:C) and peptidoglycan (PGN). Immunofluorescence microscopy and Western blotting analyses revealed that gcPGRP6 is effectively secreted to the exterior of CIK cells. The over-expression of gcPGRP6 in CIK cells leads to the activation of NF-kappa B and the inhibition of intracellular bacterial growth. Moreover, cell lysates from CIK cells transfected with pTurbo-gcPGRP6-GFP plasmid display the binding activity towards Lys-type PGN from Staphylococcus aureus and DAP-type PGN from Bacillus subtilis. Furthermore, proinflammatory cytokine IL-2 and intracellular PGN receptor NOD2 had a significantly increased expression in CIK cells overexpressed with gcPGRP6. It is demonstrated that the PGRP6 in grass carp has a role in binding PGN, in inhibiting the growth of intracellular bacteria, and in activating NF-kappa B, as well as in regulating innate immune genes. (C) 2013 Elsevier Ltd. All rights reserved.
英文摘要	Peptidoglycan recognition proteins (PGRPs) are pattern recognition molecules of innate immunity. In this study, a long-form PGRP, designated as gcPGRP6, was identified from grass carp Ctenopharyngodon idella. The deduced amino acid sequence of gcPGRP6 is composed of 464 residues with a conserved PGRP domain at the C-terminus. The gcPGRP6 gene consists of four exons and three introns, spacing approximately 2.7 kb of genomic sequence. Phylogenetic analysis demonstrated that gcPGRP6 is clustered closely with zebrafish PGLYRP6, and formed a long-type PGRP subfamily together with PGLYRP2 members identified in teleosts and mammals. Real-time PCR and Western blotting analyses revealed that gcPGRP6 is constitutively expressed in organs/tissues examined, and its expression was significantly induced in liver and intestine of grass carp in response to PGN stimulation and in CIK cells treated with lipoteichoic acid (LTA), polyinosinic polycytidylic acid (Poly I:C) and peptidoglycan (PGN). Immunofluorescence microscopy and Western blotting analyses revealed that gcPGRP6 is effectively secreted to the exterior of CIK cells. The over-expression of gcPGRP6 in CIK cells leads to the activation of NF-kappa B and the inhibition of intracellular bacterial growth. Moreover, cell lysates from CIK cells transfected with pTurbo-gcPGRP6-GFP plasmid display the binding activity towards Lys-type PGN from Staphylococcus aureus and DAP-type PGN from Bacillus subtilis. Furthermore, proinflammatory cytokine IL-2 and intracellular PGN receptor NOD2 had a significantly increased expression in CIK cells overexpressed with gcPGRP6. It is demonstrated that the PGRP6 in grass carp has a role in binding PGN, in inhibiting the growth of intracellular bacteria, and in activating NF-kappa B, as well as in regulating innate immune genes. (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 ; INNATE IMMUNITY ; DROSOPHILA-MELANOGASTER ; CRYSTAL-STRUCTURE ; EPITHELIAL-CELLS ; TELEOST FISH ; EXPRESSION ; ACTIVATION ; TOLL ; RECEPTORS
收录类别	SCI
资助信息	National Basic Research Programme of China (973 Programme) [2009CB118703]; National Natural Science Foundation of China [31172408]
语种	英语
WOS记录号	WOS:000328725800014
公开日期	2014-08-08
内容类型	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/342005/20058]
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, Hubei Province, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100039, Peoples R China
推荐引用方式 GB/T 7714	Li, Jun Hua,Yu, Zhang Long,Xue, Na Na,et al. Molecular cloning and functional characterization of peptidoglycan recognition protein 6 in grass carp Ctenopharyngodon idella[J]. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,2014,42(2):244-255.
APA	Li, Jun Hua.,Yu, Zhang Long.,Xue, Na Na.,Zou, Peng Fei.,Hu, Jing Yu.,...&Chang, Ming Xian.(2014).Molecular cloning and functional characterization of peptidoglycan recognition protein 6 in grass carp Ctenopharyngodon idella.DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,42(2),244-255.
MLA	Li, Jun Hua,et al."Molecular cloning and functional characterization of peptidoglycan recognition protein 6 in grass carp Ctenopharyngodon idella".DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 42.2(2014):244-255.