Functional analyses of the human metallothionein-IG gene. In vitro and in vivo studies
We have analyzed the human (h) metallothionein (MT)-IG proximal promoter region (-174 to +5) using a TATA box mutation (TATCA) and four trinucleotide mutants of the proximal MREa. Transient transfection of HepG2 cells was complemented by in vitro transcription with rat liver nuclear extracts. In bot...
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| Veröffentlicht in: | The Journal of biological chemistry 1995-10, Vol.270 (42), p.25194-25199 |
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| container_issue | 42 |
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| container_title | The Journal of biological chemistry |
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| creator | Samson, S L Paramchuk, W J Shworak, N W Gedamu, L |
| description | We have analyzed the human (h) metallothionein (MT)-IG proximal promoter region (-174 to +5) using a TATA box mutation (TATCA) and four trinucleotide mutants of the proximal MREa. Transient transfection of HepG2 cells was complemented by in vitro transcription with rat liver nuclear extracts. In both systems, mutations of the TATA box and conserved core of metal responsive element (MRE)a were detrimental to hMT-IG promoter activity suggesting that both elements make significant contributions to hMT-IG transcription. Although MRE binding factors were active in vitro, further metal activation of MT promoter activity was accomplished only by in vivo metal treatment rather than addition of zinc in vitro. Southwestern blotting identified nuclear proteins in rat liver and HepG2 cells which physically interact with MREa in a zinc-dependent manner and could be responsible for MREa function in each system. In addition, the functional effects of the TATCA mutation correlate with altered physical interaction with TATA box-binding protein as observed using DNase I protection. |
| doi_str_mv | 10.1074/jbc.270.42.25194 |
| format | Article |
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Although MRE binding factors were active in vitro, further metal activation of MT promoter activity was accomplished only by in vivo metal treatment rather than addition of zinc in vitro. Southwestern blotting identified nuclear proteins in rat liver and HepG2 cells which physically interact with MREa in a zinc-dependent manner and could be responsible for MREa function in each system. 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In vitro and in vivo studies</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>We have analyzed the human (h) metallothionein (MT)-IG proximal promoter region (-174 to +5) using a TATA box mutation (TATCA) and four trinucleotide mutants of the proximal MREa. Transient transfection of HepG2 cells was complemented by in vitro transcription with rat liver nuclear extracts. In both systems, mutations of the TATA box and conserved core of metal responsive element (MRE)a were detrimental to hMT-IG promoter activity suggesting that both elements make significant contributions to hMT-IG transcription. Although MRE binding factors were active in vitro, further metal activation of MT promoter activity was accomplished only by in vivo metal treatment rather than addition of zinc in vitro. Southwestern blotting identified nuclear proteins in rat liver and HepG2 cells which physically interact with MREa in a zinc-dependent manner and could be responsible for MREa function in each system. In addition, the functional effects of the TATCA mutation correlate with altered physical interaction with TATA box-binding protein as observed using DNase I protection.</description><subject>Animals</subject><subject>Base Sequence</subject><subject>Deoxyribonuclease I - pharmacology</subject><subject>DNA-Binding Proteins - physiology</subject><subject>Genes, Regulator</subject><subject>Humans</subject><subject>Male</subject><subject>Metallothionein - genetics</subject><subject>Molecular Sequence Data</subject><subject>Mutation</subject><subject>Promoter Regions, Genetic</subject><subject>Rats</subject><subject>Rats, Sprague-Dawley</subject><subject>TATA Box</subject><subject>TATA-Box Binding Protein</subject><subject>Transcription Factors - physiology</subject><subject>Transcription, Genetic - drug effects</subject><subject>Tumor Cells, Cultured</subject><subject>Zinc - pharmacology</subject><issn>0021-9258</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNotkL1PwzAQxT2ASinsLEie2BL8nXhEFS2VKrEAa2Q7NnWV2CV2KvW_J1X7hnd60u9OpwfAE0YlRhV73WtTkgqVjJSEY8luwBwhggtJeH0H7lPao0lM4hmYVZxLwfkc_KzGYLKPQXVQTXZKNsHoYN5ZuBt7FWBvs-q6mHcTZH0oNmv4a4Mt4SbAo89DnPZa6M_hGGHKY-ttegC3TnXJPl7nAnyv3r-WH8X2c71Zvm2LA6FVLrSxtVSKiOkZp53jjlFCjFBCOScc0QZXjDLUaukMVtjpmkqDa11hyhyRdAFeLncPQ_wbbcpN75OxXaeCjWNqsJB1jcQZfL6Co-5t2xwG36vh1FyLoP9ai18b</recordid><startdate>19951020</startdate><enddate>19951020</enddate><creator>Samson, S L</creator><creator>Paramchuk, W J</creator><creator>Shworak, N W</creator><creator>Gedamu, L</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7TM</scope></search><sort><creationdate>19951020</creationdate><title>Functional analyses of the human metallothionein-IG gene. 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Although MRE binding factors were active in vitro, further metal activation of MT promoter activity was accomplished only by in vivo metal treatment rather than addition of zinc in vitro. Southwestern blotting identified nuclear proteins in rat liver and HepG2 cells which physically interact with MREa in a zinc-dependent manner and could be responsible for MREa function in each system. In addition, the functional effects of the TATCA mutation correlate with altered physical interaction with TATA box-binding protein as observed using DNase I protection.</abstract><cop>United States</cop><pmid>7559655</pmid><doi>10.1074/jbc.270.42.25194</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of biological chemistry, 1995-10, Vol.270 (42), p.25194-25199 |
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| language | eng |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Animals Base Sequence Deoxyribonuclease I - pharmacology DNA-Binding Proteins - physiology Genes, Regulator Humans Male Metallothionein - genetics Molecular Sequence Data Mutation Promoter Regions, Genetic Rats Rats, Sprague-Dawley TATA Box TATA-Box Binding Protein Transcription Factors - physiology Transcription, Genetic - drug effects Tumor Cells, Cultured Zinc - pharmacology |
| title | Functional analyses of the human metallothionein-IG gene. In vitro and in vivo studies |
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