Molecular and functional comparison of 1,25-dihydroxyvitamin D3 and the novel vitamin D receptor ligand, lithocholic acid, in activating transcription of cytochrome P450 3A4

Peter W. Jurutka, Paul D. Thompson, G Kerr Whitfield, Kristina R. Eichhorst, Neal Hall, Carlos Encinas Dominguez, Jui-Cheng Hsieh, Carol A. Haussler, Mark R Haussler

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

The vitamin D receptor (VDR) binds to and mediates the effects of the 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) hormone to alter gene transcription. A newly recognized VDR ligand is the carcinogenic bile acid, lithocholic acid (LCA). We demonstrate that, in HT-29 colon cancer cells, both LCA and 1,25(OH)2D3 induce expression of cytochrome P450 3A4 (CYP3A4), an enzyme involved in cellular detoxification. We also show that LCA-VDR stimulates transcription of gene reporter constructs containing DR3 and ER6 vitamin D responsive elements (VDREs) from the human CYP3A4 gene. Utilizing gel mobility shift, pulldown, and mammalian two-hybrid assays, we observe that: (i) 1,25(OH)2D3 enhances retinoid X receptor (RXR) heterodimerization with VDR more effectively than LCA, (ii) the 1,25(OH)2D3-liganded VDR-RXR heterodimer recruits full-length SRC-1 coactivator, whereas this interaction is minimal with LCA unless LXXLL-containing fragments of SRC-1 are employed, and (iii) both 1,25(OH)2D3 and LCA enhance the binding of VDR to DRIP205/mediator, but unlike 1,25(OH)2D3-VDR, LCA-VDR does not interact detectably with NCoA-62 or TRIP1/SUG1, suggesting a different pattern of LCA-VDR comodulator association. Finally, residues in the human VDR (hVDR) ligand binding domain (LBD) were altered to create mutants unresponsive to 1,25(OH)2D3- and/or LCA-stimulated transactivation, identifying S237 and S225/S278 as critical for 1,25(OH)2D3 and LCA action, respectively. Therefore, these two VDR ligands contact distinct residues in the binding pocket, perhaps generating unique receptor conformations that determine the degree of RXR and comodulator binding. We propose that VDR is a bifunctional regulator, with the 1,25(OH) 2D3-liganded conformation facilitating high affinity endocrine actions, and the LCA-liganded configuration mediating local, lower affinity cellular detoxification by upregulation of CYP3A4 in the colon.

Original languageEnglish (US)
Pages (from-to)917-943
Number of pages27
JournalJournal of Cellular Biochemistry
Volume94
Issue number5
DOIs
StatePublished - Apr 1 2005

Fingerprint

Lithocholic Acid
Cytochrome P-450 CYP3A
Calcitriol Receptors
Calcitriol
Transcription
Ligands
Retinoid X Receptors
Detoxification
Genes
Conformations
Mediator Complex Subunit 1
Two-Hybrid System Techniques
Bile Acids and Salts
Reporter Genes
Vitamin D
Colonic Neoplasms
Cytochrome P-450 Enzyme System
Transcriptional Activation

Keywords

  • Coactivators
  • Colon cancer
  • Detoxification
  • Ligand-generated nuclear receptor conformation
  • Mediator
  • Retinoid X receptor
  • Vitamin D responsive elements

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Molecular and functional comparison of 1,25-dihydroxyvitamin D3 and the novel vitamin D receptor ligand, lithocholic acid, in activating transcription of cytochrome P450 3A4. / Jurutka, Peter W.; Thompson, Paul D.; Whitfield, G Kerr; Eichhorst, Kristina R.; Hall, Neal; Dominguez, Carlos Encinas; Hsieh, Jui-Cheng; Haussler, Carol A.; Haussler, Mark R.

In: Journal of Cellular Biochemistry, Vol. 94, No. 5, 01.04.2005, p. 917-943.

Research output: Contribution to journalArticle

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abstract = "The vitamin D receptor (VDR) binds to and mediates the effects of the 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) hormone to alter gene transcription. A newly recognized VDR ligand is the carcinogenic bile acid, lithocholic acid (LCA). We demonstrate that, in HT-29 colon cancer cells, both LCA and 1,25(OH)2D3 induce expression of cytochrome P450 3A4 (CYP3A4), an enzyme involved in cellular detoxification. We also show that LCA-VDR stimulates transcription of gene reporter constructs containing DR3 and ER6 vitamin D responsive elements (VDREs) from the human CYP3A4 gene. Utilizing gel mobility shift, pulldown, and mammalian two-hybrid assays, we observe that: (i) 1,25(OH)2D3 enhances retinoid X receptor (RXR) heterodimerization with VDR more effectively than LCA, (ii) the 1,25(OH)2D3-liganded VDR-RXR heterodimer recruits full-length SRC-1 coactivator, whereas this interaction is minimal with LCA unless LXXLL-containing fragments of SRC-1 are employed, and (iii) both 1,25(OH)2D3 and LCA enhance the binding of VDR to DRIP205/mediator, but unlike 1,25(OH)2D3-VDR, LCA-VDR does not interact detectably with NCoA-62 or TRIP1/SUG1, suggesting a different pattern of LCA-VDR comodulator association. Finally, residues in the human VDR (hVDR) ligand binding domain (LBD) were altered to create mutants unresponsive to 1,25(OH)2D3- and/or LCA-stimulated transactivation, identifying S237 and S225/S278 as critical for 1,25(OH)2D3 and LCA action, respectively. Therefore, these two VDR ligands contact distinct residues in the binding pocket, perhaps generating unique receptor conformations that determine the degree of RXR and comodulator binding. We propose that VDR is a bifunctional regulator, with the 1,25(OH) 2D3-liganded conformation facilitating high affinity endocrine actions, and the LCA-liganded configuration mediating local, lower affinity cellular detoxification by upregulation of CYP3A4 in the colon.",
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AU - Thompson, Paul D.

AU - Whitfield, G Kerr

AU - Eichhorst, Kristina R.

AU - Hall, Neal

AU - Dominguez, Carlos Encinas

AU - Hsieh, Jui-Cheng

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KW - Coactivators

KW - Colon cancer

KW - Detoxification

KW - Ligand-generated nuclear receptor conformation

KW - Mediator

KW - Retinoid X receptor

KW - Vitamin D responsive elements

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