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CpG Site-Specific Regulation of Metallothionein-1 Gene Expression
Metal-binding inducible proteins called metallothioneins (MTs) protect cells from heavy-metal toxicity. Their transcription is regulated by metal response element (MRE)-binding transcription factor-1 (MTF1), which is strongly recruited to MREs in the MT promoters, in response to Zn and Cd. Mouse Mt1...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7503544/ https://www.ncbi.nlm.nih.gov/pubmed/32824906 http://dx.doi.org/10.3390/ijms21175946 |
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author | Ogushi, Shoko Yoshida, Yuya Nakanishi, Tsuyoshi Kimura, Tomoki |
author_facet | Ogushi, Shoko Yoshida, Yuya Nakanishi, Tsuyoshi Kimura, Tomoki |
author_sort | Ogushi, Shoko |
collection | PubMed |
description | Metal-binding inducible proteins called metallothioneins (MTs) protect cells from heavy-metal toxicity. Their transcription is regulated by metal response element (MRE)-binding transcription factor-1 (MTF1), which is strongly recruited to MREs in the MT promoters, in response to Zn and Cd. Mouse Mt1 gene promoter contains 5 MREs (a–e), and MTF1 has the highest affinity to MREd. Epigenetic changes like DNA methylation might affect transcription and, therefore, the cytoprotective function of MT genes. To reveal the CpG site(s) critical for Mt1 transcription, we analyzed the methylation status of CpG dinucleotides in the Mt1 gene promoter through bisulfite sequencing in P1798 mouse lymphosarcoma cells, with high or low MT expression. We found demethylated CpG sites near MREd and MREe, in cells with high expression. Next, we compared Mt1 gene-promoter-driven Lucia luciferase gene expression in unmethylated and methylated reporter vectors. To clarify the effect of complete and partial CpG methylation, we used M.SssI (CG→(5m)CG) and HhaI (GCGC→G(5m)CGC)-methylated reporter vectors. Point mutation analysis revealed that methylation of a CpG site near MREd and MREe strongly inhibited Mt1 gene expression. Our results suggest that the methylation status of this site is important for the regulation of Mt1 gene expression. |
format | Online Article Text |
id | pubmed-7503544 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-75035442020-09-23 CpG Site-Specific Regulation of Metallothionein-1 Gene Expression Ogushi, Shoko Yoshida, Yuya Nakanishi, Tsuyoshi Kimura, Tomoki Int J Mol Sci Article Metal-binding inducible proteins called metallothioneins (MTs) protect cells from heavy-metal toxicity. Their transcription is regulated by metal response element (MRE)-binding transcription factor-1 (MTF1), which is strongly recruited to MREs in the MT promoters, in response to Zn and Cd. Mouse Mt1 gene promoter contains 5 MREs (a–e), and MTF1 has the highest affinity to MREd. Epigenetic changes like DNA methylation might affect transcription and, therefore, the cytoprotective function of MT genes. To reveal the CpG site(s) critical for Mt1 transcription, we analyzed the methylation status of CpG dinucleotides in the Mt1 gene promoter through bisulfite sequencing in P1798 mouse lymphosarcoma cells, with high or low MT expression. We found demethylated CpG sites near MREd and MREe, in cells with high expression. Next, we compared Mt1 gene-promoter-driven Lucia luciferase gene expression in unmethylated and methylated reporter vectors. To clarify the effect of complete and partial CpG methylation, we used M.SssI (CG→(5m)CG) and HhaI (GCGC→G(5m)CGC)-methylated reporter vectors. Point mutation analysis revealed that methylation of a CpG site near MREd and MREe strongly inhibited Mt1 gene expression. Our results suggest that the methylation status of this site is important for the regulation of Mt1 gene expression. MDPI 2020-08-19 /pmc/articles/PMC7503544/ /pubmed/32824906 http://dx.doi.org/10.3390/ijms21175946 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Ogushi, Shoko Yoshida, Yuya Nakanishi, Tsuyoshi Kimura, Tomoki CpG Site-Specific Regulation of Metallothionein-1 Gene Expression |
title | CpG Site-Specific Regulation of Metallothionein-1 Gene Expression |
title_full | CpG Site-Specific Regulation of Metallothionein-1 Gene Expression |
title_fullStr | CpG Site-Specific Regulation of Metallothionein-1 Gene Expression |
title_full_unstemmed | CpG Site-Specific Regulation of Metallothionein-1 Gene Expression |
title_short | CpG Site-Specific Regulation of Metallothionein-1 Gene Expression |
title_sort | cpg site-specific regulation of metallothionein-1 gene expression |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7503544/ https://www.ncbi.nlm.nih.gov/pubmed/32824906 http://dx.doi.org/10.3390/ijms21175946 |
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