Coactivation of SF-1-mediated transcription of steroidogenic enzymes by Ubc9 and PIAS1

Noriko Suda, Hirotaka Shibata, Isao Kurihara, Yayoi Ikeda, Sakiko Kobayashi, Kenichi Yokota, Ayano Takeda, Ken Nakagawa, Mototsugu Oya, Masaru Murai, William E. Rainey, Takao Saruta, Hiroshi Itoh

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Steroidogenic factor-1 (SF-1) is a nuclear orphan receptor, which is essential for adrenal development and regulation of steroidogenic enzyme expression. SF-1 is posttranslationally modified by small ubiquitin-related modifier-1 (SUMO-1), thus mostly resulting in attenuation of transcription. We investigated the role of sumoylation enzymes, Ubc9 and protein inhibitors of activated STAT1 (PIAS1), in SF-1-mediated transcription of steroidogenic enzyme genes in the adrenal cortex. Coimmunoprecipitation assays showed that both Ubc9 and PIAS1 interacted with SF-1. Transient transfection assays in adrenocortical H295R cells showed Ubc9 and PIAS1 potentiated SF-1-mediated transactivation of reporter constructs containing human CYP17, CYP11A1, and CYP11B1 but not CYP11B2 promoters. Reduction of endogenous Ubc9 and PIAS1 by introducing corresponding small interfering RNA significantly reduced endogenous CYP17, CYP11A1, and CYP11B1 mRNA levels, indicating that they normally function as coactivators of SF-1. Wild type and sumoylation-inactive mutants of Ubc9 and PIAS1 can similarly enhance the SF-1-mediated transactivation of the CYP17gene, indicating that the coactivation potency of Ubc9 and PIAS1 isindependentof sumoylation activity. Chromatin immunoprecipitation assays demonstrated that SF-1, Ubc9, and PIAS1 were recruited to an endogenous CYP17 gene promoter in the context of chromatin in vivo. Immunohistochemistry and Western blotting showed that SF-1, Ubc9, and PIAS1 were expressed in the nuclei of the human adrenal cortex. In cortisol-producing adenomas, the expression pattern of SF-1 and Ubc9 were markedly increased, whereas that of PIAS1 was decreased compared with adjacent normal adrenals. These results showed the physiological roles of Ubc9 and PIAS1 as SF-1 coactivators beyond sumoylation enzymes in adrenocortical steroidogenesis and suggested their possible pathophysiological roles in human cortisol-producing adenomas.

Original languageEnglish
Pages (from-to)2266-2277
Number of pages12
JournalEndocrinology
Volume152
Issue number6
DOIs
Publication statusPublished - 2011 Jun

Fingerprint

Steroidogenic Factor 1
Enzymes
Sumoylation
Proteins
Steroid 17-alpha-Hydroxylase
Steroid 11-beta-Hydroxylase
Cholesterol Side-Chain Cleavage Enzyme
Adrenal Cortex
Adenoma
Transcriptional Activation
Hydrocortisone
Cytochrome P-450 CYP11B2
Orphan Nuclear Receptors
Chromatin Immunoprecipitation
Mutant Proteins
Ubiquitin
Small Interfering RNA
Genes
Chromatin
Transfection

ASJC Scopus subject areas

  • Endocrinology

Cite this

Coactivation of SF-1-mediated transcription of steroidogenic enzymes by Ubc9 and PIAS1. / Suda, Noriko; Shibata, Hirotaka; Kurihara, Isao; Ikeda, Yayoi; Kobayashi, Sakiko; Yokota, Kenichi; Takeda, Ayano; Nakagawa, Ken; Oya, Mototsugu; Murai, Masaru; Rainey, William E.; Saruta, Takao; Itoh, Hiroshi.

In: Endocrinology, Vol. 152, No. 6, 06.2011, p. 2266-2277.

Research output: Contribution to journalArticle

Suda, N, Shibata, H, Kurihara, I, Ikeda, Y, Kobayashi, S, Yokota, K, Takeda, A, Nakagawa, K, Oya, M, Murai, M, Rainey, WE, Saruta, T & Itoh, H 2011, 'Coactivation of SF-1-mediated transcription of steroidogenic enzymes by Ubc9 and PIAS1', Endocrinology, vol. 152, no. 6, pp. 2266-2277. https://doi.org/10.1210/en.2010-1232
Suda, Noriko ; Shibata, Hirotaka ; Kurihara, Isao ; Ikeda, Yayoi ; Kobayashi, Sakiko ; Yokota, Kenichi ; Takeda, Ayano ; Nakagawa, Ken ; Oya, Mototsugu ; Murai, Masaru ; Rainey, William E. ; Saruta, Takao ; Itoh, Hiroshi. / Coactivation of SF-1-mediated transcription of steroidogenic enzymes by Ubc9 and PIAS1. In: Endocrinology. 2011 ; Vol. 152, No. 6. pp. 2266-2277.
@article{6c85e6fc15f3468b9d7a5cabb702be8f,
title = "Coactivation of SF-1-mediated transcription of steroidogenic enzymes by Ubc9 and PIAS1",
abstract = "Steroidogenic factor-1 (SF-1) is a nuclear orphan receptor, which is essential for adrenal development and regulation of steroidogenic enzyme expression. SF-1 is posttranslationally modified by small ubiquitin-related modifier-1 (SUMO-1), thus mostly resulting in attenuation of transcription. We investigated the role of sumoylation enzymes, Ubc9 and protein inhibitors of activated STAT1 (PIAS1), in SF-1-mediated transcription of steroidogenic enzyme genes in the adrenal cortex. Coimmunoprecipitation assays showed that both Ubc9 and PIAS1 interacted with SF-1. Transient transfection assays in adrenocortical H295R cells showed Ubc9 and PIAS1 potentiated SF-1-mediated transactivation of reporter constructs containing human CYP17, CYP11A1, and CYP11B1 but not CYP11B2 promoters. Reduction of endogenous Ubc9 and PIAS1 by introducing corresponding small interfering RNA significantly reduced endogenous CYP17, CYP11A1, and CYP11B1 mRNA levels, indicating that they normally function as coactivators of SF-1. Wild type and sumoylation-inactive mutants of Ubc9 and PIAS1 can similarly enhance the SF-1-mediated transactivation of the CYP17gene, indicating that the coactivation potency of Ubc9 and PIAS1 isindependentof sumoylation activity. Chromatin immunoprecipitation assays demonstrated that SF-1, Ubc9, and PIAS1 were recruited to an endogenous CYP17 gene promoter in the context of chromatin in vivo. Immunohistochemistry and Western blotting showed that SF-1, Ubc9, and PIAS1 were expressed in the nuclei of the human adrenal cortex. In cortisol-producing adenomas, the expression pattern of SF-1 and Ubc9 were markedly increased, whereas that of PIAS1 was decreased compared with adjacent normal adrenals. These results showed the physiological roles of Ubc9 and PIAS1 as SF-1 coactivators beyond sumoylation enzymes in adrenocortical steroidogenesis and suggested their possible pathophysiological roles in human cortisol-producing adenomas.",
author = "Noriko Suda and Hirotaka Shibata and Isao Kurihara and Yayoi Ikeda and Sakiko Kobayashi and Kenichi Yokota and Ayano Takeda and Ken Nakagawa and Mototsugu Oya and Masaru Murai and Rainey, {William E.} and Takao Saruta and Hiroshi Itoh",
year = "2011",
month = "6",
doi = "10.1210/en.2010-1232",
language = "English",
volume = "152",
pages = "2266--2277",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "6",

}

TY - JOUR

T1 - Coactivation of SF-1-mediated transcription of steroidogenic enzymes by Ubc9 and PIAS1

AU - Suda, Noriko

AU - Shibata, Hirotaka

AU - Kurihara, Isao

AU - Ikeda, Yayoi

AU - Kobayashi, Sakiko

AU - Yokota, Kenichi

AU - Takeda, Ayano

AU - Nakagawa, Ken

AU - Oya, Mototsugu

AU - Murai, Masaru

AU - Rainey, William E.

AU - Saruta, Takao

AU - Itoh, Hiroshi

PY - 2011/6

Y1 - 2011/6

N2 - Steroidogenic factor-1 (SF-1) is a nuclear orphan receptor, which is essential for adrenal development and regulation of steroidogenic enzyme expression. SF-1 is posttranslationally modified by small ubiquitin-related modifier-1 (SUMO-1), thus mostly resulting in attenuation of transcription. We investigated the role of sumoylation enzymes, Ubc9 and protein inhibitors of activated STAT1 (PIAS1), in SF-1-mediated transcription of steroidogenic enzyme genes in the adrenal cortex. Coimmunoprecipitation assays showed that both Ubc9 and PIAS1 interacted with SF-1. Transient transfection assays in adrenocortical H295R cells showed Ubc9 and PIAS1 potentiated SF-1-mediated transactivation of reporter constructs containing human CYP17, CYP11A1, and CYP11B1 but not CYP11B2 promoters. Reduction of endogenous Ubc9 and PIAS1 by introducing corresponding small interfering RNA significantly reduced endogenous CYP17, CYP11A1, and CYP11B1 mRNA levels, indicating that they normally function as coactivators of SF-1. Wild type and sumoylation-inactive mutants of Ubc9 and PIAS1 can similarly enhance the SF-1-mediated transactivation of the CYP17gene, indicating that the coactivation potency of Ubc9 and PIAS1 isindependentof sumoylation activity. Chromatin immunoprecipitation assays demonstrated that SF-1, Ubc9, and PIAS1 were recruited to an endogenous CYP17 gene promoter in the context of chromatin in vivo. Immunohistochemistry and Western blotting showed that SF-1, Ubc9, and PIAS1 were expressed in the nuclei of the human adrenal cortex. In cortisol-producing adenomas, the expression pattern of SF-1 and Ubc9 were markedly increased, whereas that of PIAS1 was decreased compared with adjacent normal adrenals. These results showed the physiological roles of Ubc9 and PIAS1 as SF-1 coactivators beyond sumoylation enzymes in adrenocortical steroidogenesis and suggested their possible pathophysiological roles in human cortisol-producing adenomas.

AB - Steroidogenic factor-1 (SF-1) is a nuclear orphan receptor, which is essential for adrenal development and regulation of steroidogenic enzyme expression. SF-1 is posttranslationally modified by small ubiquitin-related modifier-1 (SUMO-1), thus mostly resulting in attenuation of transcription. We investigated the role of sumoylation enzymes, Ubc9 and protein inhibitors of activated STAT1 (PIAS1), in SF-1-mediated transcription of steroidogenic enzyme genes in the adrenal cortex. Coimmunoprecipitation assays showed that both Ubc9 and PIAS1 interacted with SF-1. Transient transfection assays in adrenocortical H295R cells showed Ubc9 and PIAS1 potentiated SF-1-mediated transactivation of reporter constructs containing human CYP17, CYP11A1, and CYP11B1 but not CYP11B2 promoters. Reduction of endogenous Ubc9 and PIAS1 by introducing corresponding small interfering RNA significantly reduced endogenous CYP17, CYP11A1, and CYP11B1 mRNA levels, indicating that they normally function as coactivators of SF-1. Wild type and sumoylation-inactive mutants of Ubc9 and PIAS1 can similarly enhance the SF-1-mediated transactivation of the CYP17gene, indicating that the coactivation potency of Ubc9 and PIAS1 isindependentof sumoylation activity. Chromatin immunoprecipitation assays demonstrated that SF-1, Ubc9, and PIAS1 were recruited to an endogenous CYP17 gene promoter in the context of chromatin in vivo. Immunohistochemistry and Western blotting showed that SF-1, Ubc9, and PIAS1 were expressed in the nuclei of the human adrenal cortex. In cortisol-producing adenomas, the expression pattern of SF-1 and Ubc9 were markedly increased, whereas that of PIAS1 was decreased compared with adjacent normal adrenals. These results showed the physiological roles of Ubc9 and PIAS1 as SF-1 coactivators beyond sumoylation enzymes in adrenocortical steroidogenesis and suggested their possible pathophysiological roles in human cortisol-producing adenomas.

UR - http://www.scopus.com/inward/record.url?scp=79956329911&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79956329911&partnerID=8YFLogxK

U2 - 10.1210/en.2010-1232

DO - 10.1210/en.2010-1232

M3 - Article

VL - 152

SP - 2266

EP - 2277

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 6

ER -