NF-YC functions as a corepressor of agonist-bound mineralocorticoid receptor

Ayano Murai-Takeda, Hirotaka Shibata, Isao Kurihara, Sakiko Kobayashi, Kenichi Yokota, Noriko Suda, Yuko Mitsuishi, Rie Jo, Hirochika Kitagawa, Shigeaki Kato, Takao Saruta, Hiroshi Itoh

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The role of aldosterone has been implicated in the metabolic syndrome and cardiovascular diseases. The biological actions of aldosterone are mediated through mineralocorticoid receptor (MR). Nuclear receptor-mediated gene expression is regulated by dynamic and coordinated recruitment of coactivators and corepressors. To identify new coregulators of the MR, full-length MR was used as bait in yeast two-hybrid screening. Weisolated NF-YC, one of the subunits of heterotrimeric transcription factor NF-Y. Specific interaction between MRand NF-YC was confirmed by yeast two-hybrid, mammalian two-hybrid, coimmunoprecipitation assays, and fluorescence subcellular imaging. Transient transfection experiments in COS-7 cells demonstrated that NF-YC repressed MR transactivation in a hormone-sensitive manner. Moreover, reduction of NF-YC protein levels by small interfering RNA potentiated hormonal activation of endogenous target genes in stably MR-expressing cells, indicating that NF-YC functions as an agonist-dependent MR corepressor. The corepressor function of NF-YC is selective for MR, because overexpression of NF-YC did not affect transcriptional activity mediated by androgen, progesterone, or glucocorticoid receptors. Chromatin immunoprecipitation experiments showed that endogenous MR and steroid receptor coactivator-1 were recruited to an endogenous ENaC gene promoter in a largely aldosterone-dependent manner, and endogenous NF-YC was sequentially recruited to the same element. Immunohistochemistry showed that endogenous MR and NF-YC were colocalized within the mouse kidney. Although aldosterone induces interaction of the N and C termini of MR, NF-YC inhibited the N/C interaction. These findings indicate that NF-YC functions as a new corepressor of agonist-bound MR via alteration of aldosterone-induced MR conformation.

Original languageEnglish
Pages (from-to)8084-8093
Number of pages10
JournalJournal of Biological Chemistry
Volume285
Issue number11
DOIs
Publication statusPublished - 2010 Mar 12

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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