TRAIL in the mandarin fish Siniperca chuatsi: Gene and its apoptotic effect in HeLa cells | |
Gao, Y.1,2,3; Chang, M. X.1,2; Sun, B. J.1,2; Nie, P.1,2; Nie, P, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei Province, Peoples R China | |
2008 | |
Source Publication | FISH & SHELLFISH IMMUNOLOGY
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ISSN | 1050-4648 |
Volume | 24Issue:1Pages:55-66 |
Abstract | Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is one of the TNF superfamily members, participating in many biological processes including cell proliferation and apoptotic death. In this study, a TRAIL gene was cloned from a perciform fish, the mandarin fish Siniperca chuatsi, a major cultured fish in China's aquaculture, and is named as SCTRAIL for S. chuatsi TRAIL. The full-length cDNA of SCTRAIL is 1359 bp, encoding a 283-amino-acid protein. This deduced protein contains the CYS231, a 23-mer fragment of transmembrane region, a glycosylation site and a TNF family signature, all of which are conserved among TRAIL members. SCTRAIL gene consists of six exons, with five intervening introns, spaced over approximately 9 kb of genomic sequence. Southern blotting demonstrated that the SCTRAIL gene is present as a single copy in mandarin fish genome. A 620 bp promoter region obtained by genome walking contains a number of putative transcription factor binding sites, such as Oct-1, Sp-1, NF-1, RAP-1, C/EBPaLp, NF-kappa B and AP-1. The SCTRAIL is constitutively expressed in all the analyzed tissues, as revealed by RT-PCR, which is confirmed by Western blotting analysis using polyclonal antibody against bacteria-derived recombinant SCTRAIL protein. As an apoptosis-inducing ligand, the overexpression of SCTRAIL but not the mutant SCTRAIL-C203S in HeLa cells induced changes characteristic of apoptosis, including chromatin condensation, nucleus fragmentation, DNA ladder, and increase of sub-G0/G1 cells in FACS analysis. (c) 2007 Elsevier Ltd. All rights reserved.; Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is one of the TNF superfamily members, participating in many biological processes including cell proliferation and apoptotic death. In this study, a TRAIL gene was cloned from a perciform fish, the mandarin fish Siniperca chuatsi, a major cultured fish in China's aquaculture, and is named as SCTRAIL for S. chuatsi TRAIL. The full-length cDNA of SCTRAIL is 1359 bp, encoding a 283-amino-acid protein. This deduced protein contains the CYS231, a 23-mer fragment of transmembrane region, a glycosylation site and a TNF family signature, all of which are conserved among TRAIL members. SCTRAIL gene consists of six exons, with five intervening introns, spaced over approximately 9 kb of genomic sequence. Southern blotting demonstrated that the SCTRAIL gene is present as a single copy in mandarin fish genome. A 620 bp promoter region obtained by genome walking contains a number of putative transcription factor binding sites, such as Oct-1, Sp-1, NF-1, RAP-1, C/EBPaLp, NF-kappa B and AP-1. The SCTRAIL is constitutively expressed in all the analyzed tissues, as revealed by RT-PCR, which is confirmed by Western blotting analysis using polyclonal antibody against bacteria-derived recombinant SCTRAIL protein. As an apoptosis-inducing ligand, the overexpression of SCTRAIL but not the mutant SCTRAIL-C203S in HeLa cells induced changes characteristic of apoptosis, including chromatin condensation, nucleus fragmentation, DNA ladder, and increase of sub-G0/G1 cells in FACS analysis. (c) 2007 Elsevier Ltd. All rights reserved. |
Subtype | Article |
Keyword | Mandarin Fish Siniperca Chuatsi Trail Apoptosis Overexpression Hela Cells |
Department | [Gao, Y.; Chang, M. X.; Sun, B. J.; Nie, P.] Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei Province, Peoples R China; [Gao, Y.; Chang, M. X.; Sun, B. J.; Nie, P.] Chinese Acad Sci, Inst Hydrobiol, Lab Fish Dis, Wuhan 430072, Hubei Province, Peoples R China; [Gao, Y.] Chinese Acad Sci, Grad Sch, Beijing 10039, Peoples R China |
Subject Area | Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences |
DOI | 10.1016/j.fsi.2007.08.001 |
WOS Headings | Science & Technology ; Life Sciences & Biomedicine |
Indexed By | SCI |
Language | 英语 |
WOS Research Area | Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences |
WOS Subject | Fisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences |
WOS ID | WOS:000253306700007 |
WOS Keyword | NECROSIS-FACTOR-ALPHA ; MOLECULAR-CLONING ; LIGAND TRAIL ; TRANSCRIPTIONAL REGULATION ; EXPRESSION ANALYSIS ; RECEPTOR ; REGION ; CDNA ; IDENTIFICATION ; INDUCTION |
Citation statistics | |
Document Type | 期刊论文 |
Identifier | http://ir.ihb.ac.cn/handle/152342/8266 |
Collection | 期刊论文 |
Corresponding Author | Nie, P, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei Province, Peoples R China |
Affiliation | 1.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei Province, Peoples R China 2.Chinese Acad Sci, Inst Hydrobiol, Lab Fish Dis, Wuhan 430072, Hubei Province, Peoples R China 3.Chinese Acad Sci, Grad Sch, Beijing 10039, Peoples R China |
Recommended Citation GB/T 7714 | Gao, Y.,Chang, M. X.,Sun, B. J.,et al. TRAIL in the mandarin fish Siniperca chuatsi: Gene and its apoptotic effect in HeLa cells[J]. FISH & SHELLFISH IMMUNOLOGY,2008,24(1):55-66. |
APA | Gao, Y.,Chang, M. X.,Sun, B. J.,Nie, P.,&Nie, P, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei Province, Peoples R China.(2008).TRAIL in the mandarin fish Siniperca chuatsi: Gene and its apoptotic effect in HeLa cells.FISH & SHELLFISH IMMUNOLOGY,24(1),55-66. |
MLA | Gao, Y.,et al."TRAIL in the mandarin fish Siniperca chuatsi: Gene and its apoptotic effect in HeLa cells".FISH & SHELLFISH IMMUNOLOGY 24.1(2008):55-66. |
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