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