Cited by 127Estrogen and related hormone effectsGenomics and Chromatin DynamicsAdrenal Hormones and Disorders

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GATA-4 and GATA-6 Modulate Tissue-Specific Transcription of the Human Gene for P450c17 by Direct Interaction with Sp1

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Flück, Christa E., Miller, Walter L.·2004·Molecular Endocrinology

DOI 10.1210/me.2003-0342

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Abstract

Cytochrome P450c17 catalyzes steroidogenic 17α-hydroxylase and 17,20 lyase activities. Expression of the gene for P450c17 is cAMP dependent, tissue specific, developmentally programmed, and varies among species. Binding of Sp1, Sp3, and NF1-C (nuclear factor 1-C) to the first 227 bp of 5′flanking DNA (−227/LUC) is crucial for basal transcription in human NCI-H295A adrenal cells. Human placental JEG-3 cells contain Sp1, Sp3, and NF1, but do not express −227/LUC, even when transfected with a vector expressing steroidogenic factor 1 (SF-1). Therefore, other factors are essential for basal expression of P450c17. Deoxyribonuclease I footprinting and EMSAs identified a GATA consensus site at −64/−58 and an SF-1 site at −58/−50. RT-PCR identified GATA-4, GATA-6, and SF-1 in NCI-H295A cells and GATA-2 and GATA-3, but not GATA-4, GATA-6, or SF-1 in JEG-3 cells. Cotransfection of either GATA-4 or GATA-6 without SF-1 activated −227/LUC in JEG-3 cells, but cotransfection of GATA-2 or GATA-3 with or without SF-1 did not. Surprisingly, mutation of the GATA binding site in −227/LUC increased GATA-4 or GATA-6 induced activity, whereas mutation of the Sp1/Sp3 site decreased it. Furthermore, promoter constructs including the GATA site, but excluding the Sp1/Sp3 site at −196/−188, were not activated by GATA-4 or GATA-6, suggesting an interaction between Sp1/Sp3 and GATA-4 or GATA-6. Glutathione-S-transferase pull-down experiments and coimmunoprecipitation demonstrated interaction between GATA-4 or GATA-6 and Sp1, but not Sp3. Chromatin immunoprecipitation assays confirmed that this GATA-4/6 interaction with Sp1 occurred at the Sp site in the P450c17 promoter in NCI-H295A cells. Demethylation with 5-aza-2-deoxycytidine permitted JEG-3 cells to express endogenous P450c17, SF-1, GATA-4, GATA-6, and transfected −227/LUC. Thus, GATA-4 or GATA-6 and Sp1 together regulate expression of P450c17 in adrenal NCI-H295A cells and methylation of P450c17, GATA-4 and GATA-6 silence the expression of P450c17 in placental JEG-3 cells.

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Authors: Flück, C. E., & Miller, W. L.
Affiliations: Department of Pediatrics and The Metabolic Research Unit, University of California San Francisco, San Francisco, California 94143-0978
University of California, San Francisco (US)
Corresponding author: Miller, Walter L.
ORCID: Miller, Walter L.
Venue: Molecular Endocrinology
Publisher: The Endocrine Society
Published: 2004-05-01
Volume: 18
Issue: 5
Pages: 1144-1157
ISSN: 0888-8809 (ISSN-L)
1944-9917
DOI: 10.1210/me.2003-0342
OpenAlex: W2147982925
Semantic Scholar: 6bff6834baa403e050cbc7e2c59f353b777e6e28
Cited by: 127
References: 71
Citations by year: 2026: 1 · 2025: 2 · 2024: 1 · 2023: 5 · 2022: 2 · 2021: 1 · 2020: 1 · 2019: 2 · 2018: 6 · 2017: 4 · 2016: 5 · 2015: 5 · 2014: 2 · 2013: 8 · 2012: 6
FWCI: 8.19
Type: journal article
Language: en
Open access: bronze
doi.org
Locations: Molecular Endocrinology (publishedVersion)
PubMed (publishedVersion)
Bern Open Repository and Information System (University of Bern) (submittedVersion)
Funders: North Carolina State University
Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung
National Institutes of Health
National Cancer Institute
Eunice Kennedy Shriver National Institute of Child Health and Human Development (HD-41958, R01HD041958)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Endocrinology › Estrogen and related hormone effects
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
Keywords: CYP17, GATA4, GATA6, Sp1, transcription factor interaction, steroidogenic factor 1, steroidogenesis, 5'-flanking region, tissue-specific gene expression, adrenal cells, placenta, transcriptional regulation, GATA binding sites
MeSH terms: Animals, Binding Sites, Cells, Cultured, Cytochrome P-450 Enzyme System, DNA-Binding Proteins, Drosophila, Female, Gene Expression Regulation, Humans, Methylation, Placenta, Promoter Regions, Genetic, Transcription Factors, Transcriptional Activation, Sp1 Transcription Factor, Neurofibromin 1, GATA4 Transcription Factor, GATA6 Transcription Factor, Steroidogenic Factor 1

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GATA-4 and GATA-6 Modulate Tissue-Specific Transcription of the Human Gene for P450c17 by Direct Interaction with Sp1

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