Cited by 46Hormonal Regulation and HypertensionEstrogen and related hormone effectsGenomics and Chromatin Dynamics

Ubc9 and protein inhibitor of activated STAT 1 activate chicken ovalbumin upstream promoter-transcription factor I-mediated human CYP11B2 gene transcription

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Kurihara, Isao, Shibata, Hirotaka, Kobayashi, Sakiko, Suda, Noriko, Ikeda, Yayoi, Yokota, Kenichi, Murai, Ayano, Saito, Ikuo +2 more·2005·Journal of Biological Chemistry

Abstract

Aldosterone synthase (CYP11B2) is involved in the final steps of aldosterone biosynthesis and expressed exclusively in the adrenal zona glomerulosa cells. Using an electrophoretic mobility shift assay, we demonstrate that COUP-TFI binds to the -129/-114 element (Ad5) of human CYP11B2 promoter. Transient transfection in H295R adrenal cells demonstrated that COUP-TFI enhanced CYP11B2 reporter activity. However, the reporter construct with mutated Ad5 sequences showed reduced basal and COUP-TFI-enhanced activity, suggesting that binding of COUP-TFI to Ad5 is important for CYP11B2 transactivation. To elucidate molecular mechanisms of COUP-TFI-mediated activity, we subsequently screened for COUP-TFI-interacting proteins from a human adrenal cDNA library using a yeast two-hybrid system and identified Ubc9 and PIAS1, which have small ubiquitin-related modifier-1 (SUMO-1) conjugase and ligase activities, respectively. The coimmunoprecipitation assays confirmed that COUP-TFI forms a complex with Ubc9 and PIAS1 in mammalian cells. Immunohistochemistry showed that Ubc9 and PIAS1 are markedly expressed in rat adrenal glomerulosa cells. Coexpression of Ubc9 and PIAS1 synergistically enhanced the COUP-TFI-mediated CYP11B2 reporter activity, indicating that both proteins function as coactivators of COUP-TFI. However, sumoylation-defective mutants, Ubc9 (C93S) and PIAS1 (C351S), continued to function as coactivators of COUP-TFI, indicating that sumoylation activity are separable from coactivator ability. In addition, chromatin immunoprecipitation assays demonstrated that ectopically expressed COUP-TFI, Ubc9, and PIAS1 were recruited to an endogenous CYP11B2 promoter. Moreover, reduction of Ubc9 or PIAS1 protein levels by small interfering RNA inhibited the CYP11B2 transactivation by COUP-TFI. Our data support a physiological role of Ubc9 and PIAS1 as transcriptional coactivators in COUP-TFI-mediated CYP11B2 transcription.

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Crossref · 10.1074/jbc.m411820200

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  • Transcriptional regulation of CYP11B2 is influenced by COUP-TF1, which is coactivated by SUMO-1 conjugase and ligase Ubc9 and PIAS1.

    “Transcriptional regulation of CYP11B2 is also influenced by the activity of chicken ovalbumin upstream promoter transcription factor I (COUP-TF1), which itself is coactivated by the small ubiquitin-related modifier-1 (SUMO-1) conjugase and ligase Ubc9 and PIAS1 (79).”

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Authors
Kurihara, I., Shibata, H., Kobayashi, S., Suda, N., Ikeda, Y., Yokota, K., Murai, A., Saito, I., Rainey, W. E., & Saruta, T.
Affiliations
  • Keio University (JP)
  • The University of Texas Southwestern Medical Center (US)
Corresponding author
Shibata, Hirotaka
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2005-02
Volume
280
Issue
8
Pages
6721-6730
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2161366240
Cited by
46
References
62
Citations by year
2024: 1 · 2023: 1 · 2022: 2 · 2021: 1 · 2019: 1 · 2017: 7 · 2016: 1 · 2015: 4 · 2014: 1 · 2013: 1 · 2012: 4
FWCI
1.48
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health (DK043140)
  • Japan Society for the Promotion of Science (14571072)
  • National Institute of Diabetes and Digestive and Kidney Diseases (DK 043140)
Topics
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Hormonal Regulation and Hypertension
  • 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
Ubc9, PIAS1, COUP-TFI, CYP11B2, aldosterone synthase, SUMOylation, transcriptional coactivator, aldosterone biosynthesis, adrenal zona glomerulosa, promoter transactivation, protein-protein interaction
MeSH terms
Ubiquitin-Conjugating Enzyme UBC9, Animals, Cell Line, DNA-Binding Proteins, Enhancer Elements, Genetic, Humans, Promoter Regions, Genetic, Proteins, Transcription Factors, Transcription, Genetic, Transfection, Zona Glomerulosa, Transcriptional Activation, Trans-Activators, Cytochrome P-450 CYP11B2, Two-Hybrid System Techniques, Small Ubiquitin-Related Modifier Proteins, Ubiquitin-Conjugating Enzymes, Protein Inhibitors of Activated STAT, Rats, COUP Transcription Factor I

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