Interaction between the retinoid X receptor and transcription factor IIB is ligand-dependent in vivo

Gary M. Leong, Ken S. Wang, Matthew J. Marton, Jorge C G Blanco, I. Ming Wang, Ronda J. Rolfes, Keiko Ozato, James Segars

Research output: Contribution to journalArticle

Abstract

The retinoid X receptor (RXR) influences gene activation through heterodimeric and homodimeric association with DNA and associates with TATA binding protein, TAF110, and cAMP response element-binding protein-binding protein; yet the molecular mechanisms responsible for gene activation by RXRs remain incompletely defined. Since the general transcription factor IIB (TFIIB) is a common target of sequence-specific transcriptional activators, we suspected that RXR might regulate target genes via an interaction with TFIIB. Coimmunoprecipitation, far Western analysis, and glutathione S- transferase binding studies indicated that murine RXRβ (mRXRβ) was capable of binding to human TFIIB in vitro. Functional analysis with a dual-hybrid yeast system and cotransfection assays revealed the interaction of mRXRβ with TFIIB to be ligand-dependent in vivo. Truncation experiments mapped the essential binding regions to the carboxyl region of mRXRβ (amino acids (aa) 254-389) and two regions in the carboxyl region of TFIIB (aa 178-201 and aa 238-271). Furthermore, the Δ390-410 mRXRβ mutant bound to TFIIB in vitro but was not active in the dual-hybrid yeast system, suggesting that the extreme carboxyl region of RXR was required for in vivo interaction with TFIIB. These data indicate that interaction of mRXRβ with TFIIB is specific, direct, and ligand-dependent in vivo and suggest that gene activation by RXR involves TFIIB.

Original languageEnglish (US)
Pages (from-to)2296-2305
Number of pages10
JournalJournal of Biological Chemistry
Volume273
Issue number4
DOIs
StatePublished - Jan 23 1998
Externally publishedYes

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Transcription Factor TFIIB
Retinoid X Receptors
Ligands
Genes
Transcriptional Activation
Chemical activation
Amino Acids
Yeast
Yeasts
General Transcription Factors
TATA-Box Binding Protein
hexachlorocyclohexane x-factor
Cyclic AMP Response Element-Binding Protein
Functional analysis
Glutathione Transferase
Protein Binding
Assays
Carrier Proteins
Association reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Interaction between the retinoid X receptor and transcription factor IIB is ligand-dependent in vivo. / Leong, Gary M.; Wang, Ken S.; Marton, Matthew J.; Blanco, Jorge C G; Wang, I. Ming; Rolfes, Ronda J.; Ozato, Keiko; Segars, James.

In: Journal of Biological Chemistry, Vol. 273, No. 4, 23.01.1998, p. 2296-2305.

Research output: Contribution to journalArticle

Leong, GM, Wang, KS, Marton, MJ, Blanco, JCG, Wang, IM, Rolfes, RJ, Ozato, K & Segars, J 1998, 'Interaction between the retinoid X receptor and transcription factor IIB is ligand-dependent in vivo', Journal of Biological Chemistry, vol. 273, no. 4, pp. 2296-2305. https://doi.org/10.1074/jbc.273.4.2296
Leong, Gary M. ; Wang, Ken S. ; Marton, Matthew J. ; Blanco, Jorge C G ; Wang, I. Ming ; Rolfes, Ronda J. ; Ozato, Keiko ; Segars, James. / Interaction between the retinoid X receptor and transcription factor IIB is ligand-dependent in vivo. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 4. pp. 2296-2305.
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