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题名: Molecular cloning and functional characterization of peptidoglycan recognition protein 6 in grass carp Ctenopharyngodon idella
作者: Li, Jun Hua1, 2; Yu, Zhang Long1, 2; Xue, Na Na1, 2; Zou, Peng Fei1, 2; Hu, Jing Yu1; Nie, P.1; Chang, Ming Xian1
通讯作者: Chang, MX (reprint author), Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei Province, Peoples R China.
关键词: Peptidoglycan recognition protein ; PGRP6 ; Peptidoglycan-binding activity ; Antibacterial activity ; Grass carp ; Ctenopharyngodon idella
刊名: DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
发表日期: 2014
DOI: 10.1016/j.dci.2013.09.014
卷: 42, 期:2, 页:244-255
收录类别: SCI
文章类型: Article
部门归属: [Li, Jun Hua ; Yu, Zhang Long ; Xue, Na Na ; Zou, Peng Fei ; Hu, Jing Yu ; Nie, P. ; Chang, Ming Xian] Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei Province, Peoples R China ; [Li, Jun Hua ; Yu, Zhang Long ; Xue, Na Na ; Zou, Peng Fei] Chinese Acad Sci, Grad Univ, Beijing 100039, Peoples R China
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
资助者: National Basic Research Programme of China (973 Programme) [2009CB118703]; National Natural Science Foundation of China [31172408]
类目[WOS]: Immunology ; Zoology
研究领域[WOS]: Immunology ; Zoology
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.
关键词[WOS]: NF-KAPPA-B ; INNATE IMMUNITY ; DROSOPHILA-MELANOGASTER ; CRYSTAL-STRUCTURE ; EPITHELIAL-CELLS ; TELEOST FISH ; EXPRESSION ; ACTIVATION ; TOLL ; RECEPTORS
语种: 英语
WOS记录号: WOS:000328725800014
ISSN号: 0145-305X
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/342005/20058
Appears in Collections:鱼类生物学及渔业生物技术研究中心_期刊论文

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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

Recommended Citation:
Li, Jun Hua; Yu, Zhang Long; Xue, Na Na; Zou, Peng Fei; Hu, Jing Yu; Nie, P.; Chang, Ming Xian.Molecular cloning and functional characterization of peptidoglycan recognition protein 6 in grass carp Ctenopharyngodon idella,DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,2014,42(2):244-255
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