Hormone-dependent Translocation of Vitamin D Receptors Is Linked to Transactivation

Rácz, Attila; Bársony, Julia

doi:10.1074/jbc.274.27.19352

Cited by 104 publications

(69 citation statements)

References 42 publications

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“…8a). We have detected similar intranuclear VDR-GFP foci formation after hormone treatment (5). However, the addition of calcitriol did not cause such focal nuclear accumulation of the dnaGFP-VDR and the nlsGFP-VDR (Fig.…”

Section: Fig 2 Mutation In the Nls Of Gfp-vdr Perturbs Subcellularsupporting

confidence: 51%

“…Cell Culture-ROS17/2.8 rat osteosarcoma cells (ROS), CV-1 and COS-7 African green monkey cells, and 293 human kidney epithelial cells were obtained and grown as described previously (5). Cell cultures were maintained in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (HyClone), 2 mM glutamine, 0.1 M insulin, and 0.1 mg/ml gentamicin.…”

Section: Methodsmentioning

confidence: 99%

“…We found earlier that the cytoplasmic portion of VDR-GFP also translocates into the nucleus in a ligand-dependent fashion (5,6). The mechanisms controlling VDR translocation, however, remained to be elucidated.…”

mentioning

confidence: 99%

“…Recently we and others have used green fluorescent protein chimeras of VDR to study the receptor distribution in living cells (5)(6)(7). Unlike the glucocorticoid receptor (GR), which stays in the cytoplasm without the ligand, the unliganded VDR distributes evenly between the cytoplasm and the nucleus.…”

mentioning

confidence: 99%

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Dimerization with Retinoid X Receptors Promotes Nuclear Localization and Subnuclear Targeting of Vitamin D Receptors

Prüfer

Rácz

Lin

et al. 2000

Journal of Biological Chemistry

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133

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The vitamin D receptor (VDR) acts as heterodimer with the retinoid X receptor ␣ (RXR) to control transcriptional activity of target genes. To explore the influence of heterodimerization on the subcellular distribution of these receptors in living cells, we developed a series of fluorescent-protein chimeras. The steady-state distribution of the yellow fluorescent protein-RXR was more nuclear than the unliganded green fluorescent protein (GFP)-VDR. Coexpression of RXR-blue fluorescent protein (BFP) promoted nuclear accumulation of GFP-VDR by influencing both nuclear import and retention. Fluorescence resonance energy transfer microscopy (FRET) demonstrated that the unliganded GFP-VDR and RXR-BFP form heterodimers. The increase in nuclear heterodimer content correlated with an increase in basal transcriptional activity. FRET also revealed that calcitriol induces formation of multiple nuclear foci of heterodimers. Mutational analysis showed a correlation between hormone-dependent nuclear VDR foci formation and DNA binding. RXR-BFP also promoted hormone-dependent nuclear accumulation and intranuclear foci formation of a nuclear localization signal mutant receptor (nlsGFP-VDR) and rescued its transcriptional activity. Heterodimerization mutant RXR failed to alter GFP-VDR and nlsGFP-VDR distribution or activity. These experiments suggest that RXR has a profound effect on VDR distribution. This effect of RXR to promote nuclear accumulation and intranuclear targeting contributes to the regulation of VDR activity and probably the activity of other heterodimerization partners.Proteins of the nuclear receptor superfamily mediate response to hormones or intracellular signals into transcriptional responses and regulate an array of important cellular functions. A member of the nuclear receptor superfamily, the vitamin D receptor (VDR) 1 , mediates effects of calcitriol on bone development and maintenance, calcium homeostasis, immune functions, endocrine functions, vitamin D metabolism, and cellular proliferation and differentiation. Like other class II nuclear receptors, such as the thyroid hormone receptor, the retinoic acid receptor, and many orphan receptors, VDR requires heterodimerization with the retinoid X receptor (RXR) for high affinity binding to target genes (1, 2). VDR and RXR can heterodimerize in the absence of calcitriol, and these heterodimers regulate basal transcriptional activity of target genes and exert transcriptional silencing functions (3). The addition of calcitriol stabilizes the heterodimers and promotes their binding to the vitamin D response elements (4). The importance of heterodimerization in VDR functions led us to investigate the spatial and temporal relationships between these receptors in living cells.Recently we and others have used green fluorescent protein chimeras of VDR to study the receptor distribution in living cells (5-7). Unlike the glucocorticoid receptor (GR), which stays in the cytoplasm without the ligand, the unliganded VDR distributes evenly between the cytoplasm and the nucle...

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Section: Fig 2 Mutation In the Nls Of Gfp-vdr Perturbs Subcellularsupporting

confidence: 51%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Dimerization with Retinoid X Receptors Promotes Nuclear Localization and Subnuclear Targeting of Vitamin D Receptors

Prüfer

Rácz

Lin

et al. 2000

Journal of Biological Chemistry

Self Cite

133

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“…HEK293, and F9 cells were obtained from ATCC (Manassas, VA) and grown as described previously [Racz and Barsony, 1999;Prufer et al, 2000]. Briefly, cell cultures were maintained in DMEM (Invitrogen, Carlsbad, CA) supplemented with 10% FBS (Hyclone, Logan, UT), 2 mM glutamine (Invitrogen), and 0.1 mg/ml gentamicin (Invitrogen …”

Section: Cells and Expression Constructsmentioning

confidence: 99%

Nuclear localization of liver X receptor α and β is differentially regulated

Prüfer

Boudreaux

2006

J of Cellular Biochemistry

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Activity of nuclear receptors is regulated by their nuclear localization. Liver X receptors (LXR) a and b are nuclear receptors that regulate transcription of genes for cholesterol metabolism, cholesterol transport, and lipogenesis. While LXR a and b are very similar in structure and exhibit similar ligand binding properties, their physiological roles are quite different. Since the LXRs fall into a class of receptors that move between the nucleus and cytoplasm, experiments were conducted to determine whether LXR a and LXR b show differences in their nuclear localization pattern. To determine the location of each receptor, cell lines stably expressing yellow fluorescent protein (YFP) chimeras with either LXR a or LXR b were examined. Retention in the nucleus of the chimeric proteins in the presence or absence of ligands was assessed using fluorescence microscopy coupled with digitonin permeabilization assays. Surprisingly, differences were found between LXR a and LXR b. Whereas unliganded LXR a was retained in the nucleus, unliganded LXR b was partially exported. Mutations were then introduced into putative nuclear localization sequences (NLS) to determine which sequences are important for nuclear localization and function. Mutation in one such sequence abolished nuclear localization of LXR a, whereas the analogous change in LXR b had a much less dramatic effect. Mutations in analogous putative NLS also differentially affected transcriptional activation by LXR a and LXR b. These data demonstrate for the first time that nuclear retention and localization as well as function of LXR a and LXR b are differentially regulated.

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Effect of 1,25-dihydroxyvitamin D3 on cultured mesencephalic dopaminergic neurons to the combined toxicity caused by L-buthionine sulfoximine and 1-methyl-4-phenylpyridine

et al. 2000

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A decrease in intracellular glutathione content may be related to the primary event in Parkinson's disease, so increasing the glutathione level may have a therapeutic benefit. The biologically active form of vitamin D, 1,25‐dihydroxyvitamin D3 [1,25‐(OH)2D3] has been recently reported to enhance the intracellular glutathione concentration in the central nervous system. Exposing rat cultured mesencephalic neurons for 24 hr to a mixture of L‐buthionine sulfoximine (BSO) and 1‐methyl‐4‐phenylpyridium ions (MPP+) resulted in a relatively selective damage to dopaminergic neurons. This damage has been accompanied by a reduction of intracellular glutathione levels. Low doses, i.e., 1–100 nM, of 1,25‐(OH)2D3 protect cultured dopaminergic neurons against this toxicity, although higher concentrations of this active form of vitamin D have been found to enhance the toxic effect. Generation of reactive oxygen species (ROS) by this toxicity has been attenuated in cultures being pretreated with low concentrations of 1,25‐(OH)2D3. Because the hormone increases the intracellular glutathione content in cultures, determining how this hormone suppresses ROS generation may involve the enhancement of the antioxidative system. These data suggest that low doses of 1,25‐(OH)2D3 are able to protect mesencephalic dopaminergic neurons against BSO/MPP+‐induced toxicity that causes a depletion in glutathione content. J. Neurosci. Res. 62:374–382, 2000. © 2000 Wiley‐Liss, Inc.

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Hormone-dependent Translocation of Vitamin D Receptors Is Linked to Transactivation

Cited by 104 publications

References 42 publications

Dimerization with Retinoid X Receptors Promotes Nuclear Localization and Subnuclear Targeting of Vitamin D Receptors

Dimerization with Retinoid X Receptors Promotes Nuclear Localization and Subnuclear Targeting of Vitamin D Receptors

Nuclear localization of liver X receptor α and β is differentially regulated

Effect of 1,25-dihydroxyvitamin D3 on cultured mesencephalic dopaminergic neurons to the combined toxicity caused by L-buthionine sulfoximine and 1-methyl-4-phenylpyridine

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