IHB OpenIR  > 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 Hua1,2; Yu, Zhang Long1,2; Xue, Na Na1,2; Zou, Peng Fei1,2; Hu, Jing Yu1; Nie, P.1; Chang, Ming Xian1; Chang, MX (reprint author), Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei Province, Peoples R China.
2014
Source PublicationDEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
ISSN0145-305X
Volume42Issue:2Pages:244-255
AbstractPeptidoglycan 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.; 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.
SubtypeArticle
KeywordPeptidoglycan Recognition Protein Pgrp6 Peptidoglycan-binding Activity Antibacterial Activity Grass Carp Ctenopharyngodon Idella
Department[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
DOI10.1016/j.dci.2013.09.014
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Funding OrganizationNational Basic Research Programme of China (973 Programme) [2009CB118703]; National Natural Science Foundation of China [31172408] ; National Basic Research Programme of China (973 Programme) [2009CB118703]; National Natural Science Foundation of China [31172408] ; National Basic Research Programme of China (973 Programme) [2009CB118703]; National Natural Science Foundation of China [31172408] ; National Basic Research Programme of China (973 Programme) [2009CB118703]; National Natural Science Foundation of China [31172408]
Indexed BySCI
Language英语
WOS Research AreaImmunology ; Zoology
WOS SubjectImmunology ; Zoology
WOS IDWOS:000328725800014
WOS KeywordNF-KAPPA-B ; INNATE IMMUNITY ; DROSOPHILA-MELANOGASTER ; CRYSTAL-STRUCTURE ; EPITHELIAL-CELLS ; TELEOST FISH ; EXPRESSION ; ACTIVATION ; TOLL ; RECEPTORS
Funding OrganizationNational Basic Research Programme of China (973 Programme) [2009CB118703]; National Natural Science Foundation of China [31172408] ; National Basic Research Programme of China (973 Programme) [2009CB118703]; National Natural Science Foundation of China [31172408] ; National Basic Research Programme of China (973 Programme) [2009CB118703]; National Natural Science Foundation of China [31172408] ; National Basic Research Programme of China (973 Programme) [2009CB118703]; National Natural Science Foundation of China [31172408]
Citation statistics
Cited Times:19[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/20058
Collection鱼类生物学及渔业生物技术研究中心_期刊论文
Corresponding AuthorChang, MX (reprint author), Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei Province, Peoples R China.
Affiliation1.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
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, MX .(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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