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Antimicrobial activity-specific to Gram-negative bacteria and immune modulation-mediated NF-kappa B and Sp1 of a medaka beta-defensin
Zhao, Jiu-Gang; Zhou, Li; Jin, Jun-Yan; Zhao, Zhe; Lan, Jing; Zhang, Yi-Bin; Zhang, Qi-Ya; Gui, Jian-Fang; Gui, JF, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, 7 Donghu S Rd, Wuhan 430072, Peoples R China
2009
Source PublicationDEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
ISSN0145-305X
Volume33Issue:4Pages:624-637
AbstractDefensins are a group of cationic antimicrobial peptides which play an important role in the innate immune system by exerting their antimicrobial activity against pathogens. In this study, we cloned a novel beta-defensin cDNA from medaka (Oryzias latipes) by rapid amplification of cDNA ends (RACE) technique. The full-length cDNA consists of 480 bp, and the open reading frame (CRF) of 189 bp encodes a polypeptide of 63 amino acids (aa) with a predicted molecular weight of 7.44 kDa. Its genomic organization was analyzed, and Southern blot detection confirmed that only one copy of beta-defensin exists in the medaka HNI strain. RT-PCR, Western blot and immunohistochemistry detections showed that the beta-defensin transcript and protein could be detected in eyes, liver, kidney, blood, spleen and gill, and obviously prevalent expression was found in eyes. Antimicrobial activity of the medaka beta-defensin was evaluated, and the antibacterial activity-specific to Gram-negative bacteria was revealed. Furthermore, the lipopolysaccharide (LPS), a major component of the outer membrane of Gram-negative bacteria, was demonstrated to be able to induce about 13-fol up-regulation of the beta-defensin within first 12 h. In addition, promoter and promoter mutagenesis analysis were performed in the medaka beta-defensin. A proximal 100 base pair(bp) sequence (+26 to -73)and the next 1700 bp sequence (-73 to -1755) were demonstrated to be responsible for the basal promoter activity and for the transcription regulation. Three nuclear factor kappa B (NF-kappa B) cis-elements and a Sp1 cis-element were revealed by mutagenesis analysis to exist in the 5' flanking sequence, and they were confirmed to be responsible for the up-regulation of medaka beta-defensin stimulated by LPS. And, the Sp1 cis-element was further revealed to be related to the basal promoter activity, and transcriptional factor II D (TFIID) was found to be in charge of the gene transcription initiation. All the obtained data suggested that the novel medaka beta-defensin should have antimicrobial activity-specific to Gram-negative bacteria, and the antibacterial immune function should be modulated by NF-kappa B and Sp1. (C) 2008 Elsevier Ltd. All rights reserved.; Defensins are a group of cationic antimicrobial peptides which play an important role in the innate immune system by exerting their antimicrobial activity against pathogens. In this study, we cloned a novel beta-defensin cDNA from medaka (Oryzias latipes) by rapid amplification of cDNA ends (RACE) technique. The full-length cDNA consists of 480 bp, and the open reading frame (CRF) of 189 bp encodes a polypeptide of 63 amino acids (aa) with a predicted molecular weight of 7.44 kDa. Its genomic organization was analyzed, and Southern blot detection confirmed that only one copy of beta-defensin exists in the medaka HNI strain. RT-PCR, Western blot and immunohistochemistry detections showed that the beta-defensin transcript and protein could be detected in eyes, liver, kidney, blood, spleen and gill, and obviously prevalent expression was found in eyes. Antimicrobial activity of the medaka beta-defensin was evaluated, and the antibacterial activity-specific to Gram-negative bacteria was revealed. Furthermore, the lipopolysaccharide (LPS), a major component of the outer membrane of Gram-negative bacteria, was demonstrated to be able to induce about 13-fol up-regulation of the beta-defensin within first 12 h. In addition, promoter and promoter mutagenesis analysis were performed in the medaka beta-defensin. A proximal 100 base pair(bp) sequence (+26 to -73)and the next 1700 bp sequence (-73 to -1755) were demonstrated to be responsible for the basal promoter activity and for the transcription regulation. Three nuclear factor kappa B (NF-kappa B) cis-elements and a Sp1 cis-element were revealed by mutagenesis analysis to exist in the 5' flanking sequence, and they were confirmed to be responsible for the up-regulation of medaka beta-defensin stimulated by LPS. And, the Sp1 cis-element was further revealed to be related to the basal promoter activity, and transcriptional factor II D (TFIID) was found to be in charge of the gene transcription initiation. All the obtained data suggested that the novel medaka beta-defensin should have antimicrobial activity-specific to Gram-negative bacteria, and the antibacterial immune function should be modulated by NF-kappa B and Sp1. (C) 2008 Elsevier Ltd. All rights reserved.
SubtypeArticle
KeywordMedaka Beta-defensin Antibacterial Activity Promoter Analysis Nf-kappa b Sp1 Immune Modulation
Department[Zhao, Jiu-Gang; Zhou, Li; Jin, Jun-Yan; Zhao, Zhe; Lan, Jing; Zhang, Yi-Bin; Zhang, Qi-Ya; Gui, Jian-Fang] Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Subject AreaImmunology ; Zoology
DOI10.1016/j.dci.2008.11.006
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Funding OrganizationNational Major Basic Research Program [2004CB117403]; National 863 High-Technology Research Program [2006AA09Z439]; National Natural Science Foundation of China [U0631007, 30470935] ; National Major Basic Research Program [2004CB117403]; National 863 High-Technology Research Program [2006AA09Z439]; National Natural Science Foundation of China [U0631007, 30470935] ; National Major Basic Research Program [2004CB117403]; National 863 High-Technology Research Program [2006AA09Z439]; National Natural Science Foundation of China [U0631007, 30470935] ; National Major Basic Research Program [2004CB117403]; National 863 High-Technology Research Program [2006AA09Z439]; National Natural Science Foundation of China [U0631007, 30470935]
Indexed BySCI
Language英语
WOS Research AreaImmunology ; Zoology
WOS SubjectImmunology ; Zoology
WOS IDWOS:000263781100024
WOS KeywordOCULAR SURFACE TISSUE ; INNATE IMMUNITY ; EPITHELIAL-CELLS ; GENE-EXPRESSION ; TRANSCRIPTIONAL REGULATION ; DENDRITIC CELLS ; ALPHA-DEFENSINS ; SEX REVERSAL ; PEPTIDE ; FISH
Funding OrganizationNational Major Basic Research Program [2004CB117403]; National 863 High-Technology Research Program [2006AA09Z439]; National Natural Science Foundation of China [U0631007, 30470935] ; National Major Basic Research Program [2004CB117403]; National 863 High-Technology Research Program [2006AA09Z439]; National Natural Science Foundation of China [U0631007, 30470935] ; National Major Basic Research Program [2004CB117403]; National 863 High-Technology Research Program [2006AA09Z439]; National Natural Science Foundation of China [U0631007, 30470935] ; National Major Basic Research Program [2004CB117403]; National 863 High-Technology Research Program [2006AA09Z439]; National Natural Science Foundation of China [U0631007, 30470935]
Citation statistics
Cited Times:47[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/7852
Collection期刊论文
Corresponding AuthorGui, JF, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, 7 Donghu S Rd, Wuhan 430072, Peoples R China
AffiliationChinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Recommended Citation
GB/T 7714
Zhao, Jiu-Gang,Zhou, Li,Jin, Jun-Yan,et al. Antimicrobial activity-specific to Gram-negative bacteria and immune modulation-mediated NF-kappa B and Sp1 of a medaka beta-defensin[J]. DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,2009,33(4):624-637.
APA Zhao, Jiu-Gang.,Zhou, Li.,Jin, Jun-Yan.,Zhao, Zhe.,Lan, Jing.,...&Gui, JF, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, 7 Donghu S Rd, Wuhan 430072, Peoples R China.(2009).Antimicrobial activity-specific to Gram-negative bacteria and immune modulation-mediated NF-kappa B and Sp1 of a medaka beta-defensin.DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY,33(4),624-637.
MLA Zhao, Jiu-Gang,et al."Antimicrobial activity-specific to Gram-negative bacteria and immune modulation-mediated NF-kappa B and Sp1 of a medaka beta-defensin".DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY 33.4(2009):624-637.
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