Transcription factor CgMTF-1 regulates CgZnT1 and CgMT expression in Pacific oyster (Crassostrea gigas) under zinc stress
Oysters accumulate zinc at high tissue concentrations, and the metal response element (MRE)-binding transcription factor (MTF) functions as the cellular zinc sensor that coordinates the expression of genes involved in zinc efflux and storage, as well as those that protect against metal toxicity. In...
Published in: | Aquatic Toxicology |
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Online Access: | http://ir.qdio.ac.cn/handle/337002/49785 https://doi.org/10.1016/j.aquatox.2015.05.023 |
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ftchinacasciocas:oai:ir.qdio.ac.cn:337002/49785 2023-05-15T15:59:03+02:00 Transcription factor CgMTF-1 regulates CgZnT1 and CgMT expression in Pacific oyster (Crassostrea gigas) under zinc stress Meng, Jie Zhang, Linlin Li, Li Li, Chunyan Wang, Ting Zhang, Guofan 2015-08-01 http://ir.qdio.ac.cn/handle/337002/49785 https://doi.org/10.1016/j.aquatox.2015.05.023 英语 eng AQUATIC TOXICOLOGY Meng, Jie,Zhang, Linlin,Li, Li,et al. Transcription factor CgMTF-1 regulates CgZnT1 and CgMT expression in Pacific oyster (Crassostrea gigas) under zinc stress[J]. AQUATIC TOXICOLOGY,2015,165(2015):179-188. http://ir.qdio.ac.cn/handle/337002/49785 doi:10.1016/j.aquatox.2015.05.023 Crassostrea Zinc Accumulation Transcription Mechanism Zinc Homeostasis Article 期刊论文 2015 ftchinacasciocas https://doi.org/10.1016/j.aquatox.2015.05.023 2022-06-27T05:37:17Z Oysters accumulate zinc at high tissue concentrations, and the metal response element (MRE)-binding transcription factor (MTF) functions as the cellular zinc sensor that coordinates the expression of genes involved in zinc efflux and storage, as well as those that protect against metal toxicity. In this study, we cloned MTF-1 in oysters and examined its regulation mechanism for its classic target genes, including MTs and ZnT1 under zinc exposure conditions. We cloned CgMTF-1 and determined the subcellular locations of its protein product in HEK293 cells. CgMTF-1 has a 2826 bp open reading frame that encodes a predicted polypeptide with 707 amino acid residues, showing six well-conserved zinc finger domains that are required for metal binding. In HEK293 cell lines, CgMTF-1 primarily localizes in the cell nucleus under unstressed conditions and nuclear translocation was not critical for the activation of this gene. We searched for CgMTF-1-regulated genes in oysters using RNA interference. Decreased expression levels of CgMT1, CgMT4, and CgZnT1 were observed after CgMTF-1 interference (>70% inhibition) under zinc exposure, indicating the critical role of CgMTF-1 in the regulation of these genes. We searched for a direct regulation mechanism involving CgMTF-1 for CgMT1, CgMT4, and CgZnT1 in vitro. EMSA experiments indicated that CgMTF-1 can bind with the MREs found in the CgZnT1, CgMT1 and CgMT4 promoter regions. Additionally, luciferase reporter gene experiments indicated that CgMTF-1 could activate the CgMT1,CgMT4, and CgZnT1 promoters. Overall, our results suggest that CgMTF-1 directly coordinates the regulation of CgMTs and CgZnT1 expression and plays important roles in protecting oysters under zinc exposure conditions. To our knowledge, this is the first study to elucidate the function of MTF-1 in marine bivalves and provides new insights into the mechanisms of zinc accumulation and tolerance in mollusks. (C) 2015 Elsevier B.V. All rights reserved. Article in Journal/Newspaper Crassostrea gigas Pacific oyster Institute of Oceanology, Chinese Academy of Sciences: IOCAS-IR Pacific Aquatic Toxicology 165 179 188 |
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Open Polar |
collection |
Institute of Oceanology, Chinese Academy of Sciences: IOCAS-IR |
op_collection_id |
ftchinacasciocas |
language |
English |
topic |
Crassostrea Zinc Accumulation Transcription Mechanism Zinc Homeostasis |
spellingShingle |
Crassostrea Zinc Accumulation Transcription Mechanism Zinc Homeostasis Meng, Jie Zhang, Linlin Li, Li Li, Chunyan Wang, Ting Zhang, Guofan Transcription factor CgMTF-1 regulates CgZnT1 and CgMT expression in Pacific oyster (Crassostrea gigas) under zinc stress |
topic_facet |
Crassostrea Zinc Accumulation Transcription Mechanism Zinc Homeostasis |
description |
Oysters accumulate zinc at high tissue concentrations, and the metal response element (MRE)-binding transcription factor (MTF) functions as the cellular zinc sensor that coordinates the expression of genes involved in zinc efflux and storage, as well as those that protect against metal toxicity. In this study, we cloned MTF-1 in oysters and examined its regulation mechanism for its classic target genes, including MTs and ZnT1 under zinc exposure conditions. We cloned CgMTF-1 and determined the subcellular locations of its protein product in HEK293 cells. CgMTF-1 has a 2826 bp open reading frame that encodes a predicted polypeptide with 707 amino acid residues, showing six well-conserved zinc finger domains that are required for metal binding. In HEK293 cell lines, CgMTF-1 primarily localizes in the cell nucleus under unstressed conditions and nuclear translocation was not critical for the activation of this gene. We searched for CgMTF-1-regulated genes in oysters using RNA interference. Decreased expression levels of CgMT1, CgMT4, and CgZnT1 were observed after CgMTF-1 interference (>70% inhibition) under zinc exposure, indicating the critical role of CgMTF-1 in the regulation of these genes. We searched for a direct regulation mechanism involving CgMTF-1 for CgMT1, CgMT4, and CgZnT1 in vitro. EMSA experiments indicated that CgMTF-1 can bind with the MREs found in the CgZnT1, CgMT1 and CgMT4 promoter regions. Additionally, luciferase reporter gene experiments indicated that CgMTF-1 could activate the CgMT1,CgMT4, and CgZnT1 promoters. Overall, our results suggest that CgMTF-1 directly coordinates the regulation of CgMTs and CgZnT1 expression and plays important roles in protecting oysters under zinc exposure conditions. To our knowledge, this is the first study to elucidate the function of MTF-1 in marine bivalves and provides new insights into the mechanisms of zinc accumulation and tolerance in mollusks. (C) 2015 Elsevier B.V. All rights reserved. |
format |
Article in Journal/Newspaper |
author |
Meng, Jie Zhang, Linlin Li, Li Li, Chunyan Wang, Ting Zhang, Guofan |
author_facet |
Meng, Jie Zhang, Linlin Li, Li Li, Chunyan Wang, Ting Zhang, Guofan |
author_sort |
Meng, Jie |
title |
Transcription factor CgMTF-1 regulates CgZnT1 and CgMT expression in Pacific oyster (Crassostrea gigas) under zinc stress |
title_short |
Transcription factor CgMTF-1 regulates CgZnT1 and CgMT expression in Pacific oyster (Crassostrea gigas) under zinc stress |
title_full |
Transcription factor CgMTF-1 regulates CgZnT1 and CgMT expression in Pacific oyster (Crassostrea gigas) under zinc stress |
title_fullStr |
Transcription factor CgMTF-1 regulates CgZnT1 and CgMT expression in Pacific oyster (Crassostrea gigas) under zinc stress |
title_full_unstemmed |
Transcription factor CgMTF-1 regulates CgZnT1 and CgMT expression in Pacific oyster (Crassostrea gigas) under zinc stress |
title_sort |
transcription factor cgmtf-1 regulates cgznt1 and cgmt expression in pacific oyster (crassostrea gigas) under zinc stress |
publishDate |
2015 |
url |
http://ir.qdio.ac.cn/handle/337002/49785 https://doi.org/10.1016/j.aquatox.2015.05.023 |
geographic |
Pacific |
geographic_facet |
Pacific |
genre |
Crassostrea gigas Pacific oyster |
genre_facet |
Crassostrea gigas Pacific oyster |
op_relation |
AQUATIC TOXICOLOGY Meng, Jie,Zhang, Linlin,Li, Li,et al. Transcription factor CgMTF-1 regulates CgZnT1 and CgMT expression in Pacific oyster (Crassostrea gigas) under zinc stress[J]. AQUATIC TOXICOLOGY,2015,165(2015):179-188. http://ir.qdio.ac.cn/handle/337002/49785 doi:10.1016/j.aquatox.2015.05.023 |
op_doi |
https://doi.org/10.1016/j.aquatox.2015.05.023 |
container_title |
Aquatic Toxicology |
container_volume |
165 |
container_start_page |
179 |
op_container_end_page |
188 |
_version_ |
1766394824333721600 |