Abstract
We examined the role of the extracellular signal regulated kinases (ERK) in 1,25-dihydroxyvitamin D (1,25(OH)(2)D(3))-induced gene expression in the differentiated Caco-2 cells. 1,25(OH)(2)D(3)-regulated expression of the 25-hydroxyvitamin D, 24-hydroxylase (CYP24) gene (both natural gene and promoter construct) was strongly modulated by altering ERK activity (i.e., reduced by MEK inhibitors and dominant negative (dn) ERK1 and ERK2, activated by epidermal growth factor) but ERK inhibition had no effect on 1,25(OH)(2)D(3)-regulated expression of the transient receptor potential cation channel, subfamily V, member 6 (TRPV6). ERK5-mediated phosphorylation of the transcription factor Ets-1 enhanced 1,25(OH)(2)D(3)-mediated CYP24 gene transcription in proliferating but not differentiated Caco-2 cells due to reduced levels of ERK5 and Ets-1 (total and phosphoprotein levels) in differentiated cells. MEK inhibition reduced 1,25(OH)(2)D(3)-induced 3X-VDRE promoter activity but had no impact on the association of vitamin D receptor (VDR) with chromatin suggesting a role for co-activator recruitment in ERK-modulation of vitamin D-regulated CYP24 gene activation. Chromatin immunoprecipitation assays revealed that the ERK1/2 target, mediator 1 (MED1), is recruited to the CYP24, but not the TRPV6, promoter following 1,25(OH)(2)D(3) treatment. MED1 phosphorylation was sensitive to activators and inhibitors of the ERK1/2 signaling and MED1 siRNA reduced 1,25(OH)(2)D(3)-regulated human CYP24 promoter activity. This suggests ERK1/2 signaling enhances 1,25(OH)(2)D(3) effects on the CYP24 promoter by MED1-mediated events. Our data show that there are both promoter-specific and cell stage-specific roles for the ERK signaling pathway on 1,25(OH)(2)D(3)-mediated gene induction in enterocyte-like Caco-2 cells.
(c) 2008 Wiley-Liss, Inc.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Caco-2 Cells / physiology
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Calcitriol / metabolism*
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Calcium / metabolism
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Calcium Channels / genetics
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Calcium Channels / metabolism
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Enzyme Inhibitors / metabolism
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Epidermal Growth Factor / metabolism
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Gene Expression Regulation, Enzymologic*
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Humans
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Imidazoles / metabolism
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MAP Kinase Kinase 1 / antagonists & inhibitors
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MAP Kinase Kinase 1 / genetics
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MAP Kinase Kinase 1 / metabolism
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MAP Kinase Kinase 2 / antagonists & inhibitors
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MAP Kinase Kinase 2 / genetics
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MAP Kinase Kinase 2 / metabolism
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MAP Kinase Signaling System / physiology*
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Mitogen-Activated Protein Kinase 1 / antagonists & inhibitors
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Mitogen-Activated Protein Kinase 1 / genetics
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Mitogen-Activated Protein Kinase 1 / metabolism*
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Mitogen-Activated Protein Kinase 3 / antagonists & inhibitors
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Mitogen-Activated Protein Kinase 3 / genetics
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Mitogen-Activated Protein Kinase 3 / metabolism*
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Promoter Regions, Genetic
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Proto-Oncogene Protein c-ets-1 / genetics
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Proto-Oncogene Protein c-ets-1 / metabolism
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RNA Interference
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Receptors, Calcitriol / genetics
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Receptors, Calcitriol / metabolism
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Retinoid X Receptor alpha / genetics
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Retinoid X Receptor alpha / metabolism
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Steroid Hydroxylases / antagonists & inhibitors
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Steroid Hydroxylases / genetics*
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Steroid Hydroxylases / metabolism*
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TRPV Cation Channels / genetics
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TRPV Cation Channels / metabolism
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Two-Hybrid System Techniques
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Vitamin D3 24-Hydroxylase
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Vitamins / metabolism
Substances
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Calcium Channels
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Enzyme Inhibitors
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Imidazoles
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Proto-Oncogene Protein c-ets-1
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Receptors, Calcitriol
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Retinoid X Receptor alpha
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TRPV Cation Channels
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TRPV6 protein, human
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Vitamins
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Epidermal Growth Factor
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Steroid Hydroxylases
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Vitamin D3 24-Hydroxylase
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MAP2K2 protein, human
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Mitogen-Activated Protein Kinase 1
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Mitogen-Activated Protein Kinase 3
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MAP Kinase Kinase 1
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MAP Kinase Kinase 2
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MAP2K1 protein, human
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Calcitriol
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liarozole
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Calcium