Promoter independent down-regulation of the firefly luciferase gene by T3 and T3 receptor in CV1 cells
We report that the activity of the firefly luciferase (LUC) reporter gene is down-regulated by T3 and T3 receptor (TR) in the CV1 mammalian cell line, which is widely used for studies of TR action. Repression was highly reproducible, T3 and TR dependent, promoter independent, and observed regardless...
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| Veröffentlicht in: | Molecular and cellular endocrinology 1993-09, Vol.95 (1-2), p.101-109 |
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| container_title | Molecular and cellular endocrinology |
| container_volume | 95 |
| creator | TILLMAN, J. B CRONE, D. E HYOUNG-SOON KIM SPRUNG, C. N SPINDLER, S. R |
| description | We report that the activity of the firefly luciferase (LUC) reporter gene is down-regulated by T3 and T3 receptor (TR) in the CV1 mammalian cell line, which is widely used for studies of TR action. Repression was highly reproducible, T3 and TR dependent, promoter independent, and observed regardless of whether an internal control for transfection efficiency was used. Cotransfections with normal and mutant TRs indicate that the negative T3 response is mediated by sequences within the LUC gene coding region, and is not due to the interaction of TR with a limiting transcription factor. Negative regulation of the LUC reporter was overcome by a strong, cis-linked T3 response element (TRE), but continued in the presence of a TRE of moderate strength. The results described here demonstrate that conclusions drawn from studies of TRE structure and activity performed using the LUC reporter in CV1 cells should be interpreted with caution. |
| doi_str_mv | 10.1016/0303-7207(93)90034-h |
| format | Article |
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The results described here demonstrate that conclusions drawn from studies of TRE structure and activity performed using the LUC reporter in CV1 cells should be interpreted with caution.</description><identifier>ISSN: 0303-7207</identifier><identifier>EISSN: 1872-8057</identifier><identifier>DOI: 10.1016/0303-7207(93)90034-h</identifier><identifier>PMID: 8243799</identifier><identifier>CODEN: MCEND6</identifier><language>eng</language><publisher>Shannon: Elsevier</publisher><subject>Animals ; Base Sequence ; Biological and medical sciences ; Cell Line ; Cercopithecus aethiops ; Coleoptera - genetics ; Enzyme Induction - drug effects ; Fibroblasts - drug effects ; Fibroblasts - metabolism ; Fundamental and applied biological sciences. 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B</creatorcontrib><creatorcontrib>CRONE, D. E</creatorcontrib><creatorcontrib>HYOUNG-SOON KIM</creatorcontrib><creatorcontrib>SPRUNG, C. N</creatorcontrib><creatorcontrib>SPINDLER, S. R</creatorcontrib><title>Promoter independent down-regulation of the firefly luciferase gene by T3 and T3 receptor in CV1 cells</title><title>Molecular and cellular endocrinology</title><addtitle>Mol Cell Endocrinol</addtitle><description>We report that the activity of the firefly luciferase (LUC) reporter gene is down-regulated by T3 and T3 receptor (TR) in the CV1 mammalian cell line, which is widely used for studies of TR action. Repression was highly reproducible, T3 and TR dependent, promoter independent, and observed regardless of whether an internal control for transfection efficiency was used. Cotransfections with normal and mutant TRs indicate that the negative T3 response is mediated by sequences within the LUC gene coding region, and is not due to the interaction of TR with a limiting transcription factor. Negative regulation of the LUC reporter was overcome by a strong, cis-linked T3 response element (TRE), but continued in the presence of a TRE of moderate strength. The results described here demonstrate that conclusions drawn from studies of TRE structure and activity performed using the LUC reporter in CV1 cells should be interpreted with caution.</description><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Cell Line</subject><subject>Cercopithecus aethiops</subject><subject>Coleoptera - genetics</subject><subject>Enzyme Induction - drug effects</subject><subject>Fibroblasts - drug effects</subject><subject>Fibroblasts - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene expression</subject><subject>Gene Expression Regulation - drug effects</subject><subject>Genetic Vectors</subject><subject>Kidney</subject><subject>Luciferases - biosynthesis</subject><subject>Luciferases - genetics</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Molecular Sequence Data</subject><subject>Receptors, Thyroid Hormone - physiology</subject><subject>Recombinant Fusion Proteins - biosynthesis</subject><subject>Recombinant Fusion Proteins - genetics</subject><subject>Regulatory Sequences, Nucleic Acid</subject><subject>Thymidine Kinase - biosynthesis</subject><subject>Thymidine Kinase - genetics</subject><subject>Transcription Factors - metabolism</subject><subject>Transfection</subject><subject>Triiodothyronine - pharmacology</subject><issn>0303-7207</issn><issn>1872-8057</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1993</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kM1OwzAQhC0EKqXwBiD5gBAcAv5JYueIKqBIleBQuFqOvW6D0rjYiVDfngSiXnYOOzPa_RC6pOSeEpo_EE54IhgRtwW_KwjhabI5QlMqBUskycQxmh4sp-gsxi9CiMiYnKCJZCkXRTFF7j34rW8h4KqxsIN-NC22_qdJAqy7WreVb7B3uN0AdlUAV-9x3ZnKQdAR8BoawOUerzjWjR0kgIFd64dCPP-k2EBdx3N04nQd4WLUGfp4flrNF8ny7eV1_rhMDKesTSizlltrMmEl1UVuDGRAmXCyKAuac6ppmhlNpAApTVoSJlLLmBautFlWOj5DN_-9u-C_O4it2lZxuEA34LuoRE4E4ZL3xvTfaIKPsX9L7UK11WGvKFEDXjWwUwM7VXD1h1ct-tjV2N-VW7CH0Miz31-Pex2Nrl3QjaniwcZFlgsu-C9rJIJ4</recordid><startdate>199309</startdate><enddate>199309</enddate><creator>TILLMAN, J. 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Psychology</topic><topic>Gene expression</topic><topic>Gene Expression Regulation - drug effects</topic><topic>Genetic Vectors</topic><topic>Kidney</topic><topic>Luciferases - biosynthesis</topic><topic>Luciferases - genetics</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Molecular Sequence Data</topic><topic>Receptors, Thyroid Hormone - physiology</topic><topic>Recombinant Fusion Proteins - biosynthesis</topic><topic>Recombinant Fusion Proteins - genetics</topic><topic>Regulatory Sequences, Nucleic Acid</topic><topic>Thymidine Kinase - biosynthesis</topic><topic>Thymidine Kinase - genetics</topic><topic>Transcription Factors - metabolism</topic><topic>Transfection</topic><topic>Triiodothyronine - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>TILLMAN, J. B</creatorcontrib><creatorcontrib>CRONE, D. 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R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Promoter independent down-regulation of the firefly luciferase gene by T3 and T3 receptor in CV1 cells</atitle><jtitle>Molecular and cellular endocrinology</jtitle><addtitle>Mol Cell Endocrinol</addtitle><date>1993-09</date><risdate>1993</risdate><volume>95</volume><issue>1-2</issue><spage>101</spage><epage>109</epage><pages>101-109</pages><issn>0303-7207</issn><eissn>1872-8057</eissn><coden>MCEND6</coden><abstract>We report that the activity of the firefly luciferase (LUC) reporter gene is down-regulated by T3 and T3 receptor (TR) in the CV1 mammalian cell line, which is widely used for studies of TR action. Repression was highly reproducible, T3 and TR dependent, promoter independent, and observed regardless of whether an internal control for transfection efficiency was used. Cotransfections with normal and mutant TRs indicate that the negative T3 response is mediated by sequences within the LUC gene coding region, and is not due to the interaction of TR with a limiting transcription factor. Negative regulation of the LUC reporter was overcome by a strong, cis-linked T3 response element (TRE), but continued in the presence of a TRE of moderate strength. The results described here demonstrate that conclusions drawn from studies of TRE structure and activity performed using the LUC reporter in CV1 cells should be interpreted with caution.</abstract><cop>Shannon</cop><pub>Elsevier</pub><pmid>8243799</pmid><doi>10.1016/0303-7207(93)90034-h</doi><tpages>9</tpages></addata></record> |
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| issn | 0303-7207 1872-8057 |
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| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Animals Base Sequence Biological and medical sciences Cell Line Cercopithecus aethiops Coleoptera - genetics Enzyme Induction - drug effects Fibroblasts - drug effects Fibroblasts - metabolism Fundamental and applied biological sciences. Psychology Gene expression Gene Expression Regulation - drug effects Genetic Vectors Kidney Luciferases - biosynthesis Luciferases - genetics Molecular and cellular biology Molecular genetics Molecular Sequence Data Receptors, Thyroid Hormone - physiology Recombinant Fusion Proteins - biosynthesis Recombinant Fusion Proteins - genetics Regulatory Sequences, Nucleic Acid Thymidine Kinase - biosynthesis Thymidine Kinase - genetics Transcription Factors - metabolism Transfection Triiodothyronine - pharmacology |
| title | Promoter independent down-regulation of the firefly luciferase gene by T3 and T3 receptor in CV1 cells |
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