CoChat
Sign Up
Cited by 71Adrenal Hormones and DisordersEstrogen and related hormone effectsGenomics and Chromatin Dynamics

NF-1C, Sp1, and Sp3 Are Essential for Transcription of the Human Gene for P450c17 (Steroid 17α-hydroxylase/17,20 lyase) in Human Adrenal NCI-H295A Cells

Open access
Lin, Chin Jia, Martens, John W. M., Miller, Walter L.·2001·Molecular Endocrinology
DOI 10.1210/mend.15.8.0679
Open this paper in CoChat — free accountOpen PDF

Abstract

Cytochrome P450c17 catalyzes steroid 17alpha-hydroxylase and 17,20 lyase activities, which are required for the biosynthesis of cortisol and sex steroids. Human P450c17 is expressed in a cAMP-responsive, cell-specific, developmentally programmed fashion, but little is known about its transcriptional regulation. Expression of deletion mutants of up to 2,500 bp of human 5'-flanking DNA in human adrenal NCI-H295A cells indicated that most regulatory activity was confined to the first 227 bp. Deoxyribonuclease I footprinting of the proximal promoter identified the TATA box, an steroidogenic factor-1 site, and three previously uncharacterized sites at -107/85, at -178/-152, and at -220/-185. EMSAs and methylation interference assays suggested that the -107/-85 site and the -178/-152 site bind members of the NF-1 (nuclear factor-1) family of transcription factors. An NF-1 consensus sequence generated similar DNA/protein complexes, and antibodies against NF-1C2/CTF2 supershifted the complexes formed by the -107/-85 site, the -178/-152 site, and the NF-1 consensus site. Western blots of nuclear extracts from NCI-H295A cells probed with this NF-1 antiserum identified two NF-1 isoforms between 50 and 55 kDa. The presence of NF-1C2 (CTF2) and CTF5 in NCI-H295A cells was demonstrated by RT-PCR and sequencing. Mutation of both the -107/-85 and the -178/-152 NF-1 sites reduced basal transcription by half. Supershift assays showed that the ubiquitous proteins Sp1 and Sp3 both bind to the -227/-184 region, and that mutation of their binding sites reduced transcription by 75%. Mutation of the Sp1/Sp3 site plus the two NF-1 sites eliminated almost all detectable transcription. Thus, Sp1 and Sp3 binding to the -227/-184 site and NF-1C proteins binding to the -107/-85 and the -178/-152 sites are crucial for adrenal transcription of the human gene for P450c17.

Integrity signals

Registry

Crossref · 10.1210/mend.15.8.0679

Cites

15 of 15 references verified0 citation errors · 0 not checkable

Statistics

Recomputing p-values needs the paper’s full text.

Claims the paper makes

We couldn’t retrieve this paper’s full text, so its claims and self-check weren’t run. The registry and citation checks still apply.

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Cites

Flagged first, then by citations

Cited by

Sorted by citations

Authors
Lin, C. J., Martens, J. W. M., & Miller, W. L.
Affiliations
University of California, San Francisco (US)
ORCID
Venue
Molecular Endocrinology
Publisher
The Endocrine Society
Published
2001-08
Volume
15
Issue
8
Pages
1277-1293
ISSN
  • 0888-8809 (ISSN-L)
  • 1944-9917
DOI
10.1210/mend.15.8.0679
OpenAlex
W2025452617
Cited by
71
References
15
Citations by year
2024: 1 · 2020: 1 · 2017: 2 · 2016: 3 · 2015: 1 · 2014: 1 · 2013: 1 · 2012: 3
FWCI
5.31
Type
journal article
Language
en
Open access
Locations
  • Molecular Endocrinology (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • York University
  • National Institutes of Health (5r01dk042154-09)
  • School of Medicine, New York University
  • National Institute of Diabetes and Digestive and Kidney Diseases (DK-37922, DK 42154, R01DK037922)
  • Eunice Kennedy Shriver National Institute of Child Health and Human Development (HD-34449, U54HD034449)
Topics
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Keywords
transcriptional regulation, Sp1, Sp3, steroidogenic factor 1, TATA box, promoter elements, adrenal steroidogenesis, transcription factor binding
MeSH terms
Adrenal Gland Neoplasms, Adrenal Glands, Animals, Base Sequence, Binding Sites, Cell Line, Deoxyribonuclease I, DNA, DNA-Binding Proteins, HeLa Cells, Humans, Liver Neoplasms, Polymorphism, Genetic, Promoter Regions, Genetic, Steroid 17-alpha-Hydroxylase, Structure-Activity Relationship, Transcription Factors, Transcription, Genetic, Tumor Cells, Cultured, Polymerase Chain Reaction, Mutagenesis, Site-Directed, Sp1 Transcription Factor, Conserved Sequence, Sequence Analysis, DNA, DNA Footprinting, COS Cells, Mice, Sp3 Transcription Factor, NFI Transcription Factors, Steroidogenic Factor 1

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review