SIRT1 enzymatically potentiates 1,25-dihydroxyvitamin D3 signaling via vitamin D receptor deacetylation

Marya S. Sabir, Zainab Khan, Chengcheng Hu, Michael A. Galligan, Christopher M. Dussik, Sanchita Mallick, Angelika Dampf Stone, Shane F. Batie, Elizabeth T. Jacobs, G. K. Whitfield, Mark R. Haussler, Michael C. Heck, Peter Jurutka

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The hormonal metabolite of vitamin D, 1,25-dihydroxyvitamin D3 (1,25D), binds to the vitamin D receptor (VDR) and promotes heterodimerization of VDR with a retinoid-X-receptor (RXR) to genomically regulate diverse cellular processes. Herein, it is revealed for the first time that VDR is post-translationally acetylated, and that VDR immunoprecipitated from human embryonic kidney (HEK293) cells displays a dramatic decrease in acetylated receptor in the presence of 1,25D-ligand, sirtuin-1 (SIRT1) deacetylase, or the resveratrol activator of SIRT1. To elucidate the functional significance of VDR deacetylation, vitamin-d-responsive-element (VDRE)-based transcriptional assays were performed to determine if deacetylase overexpression affects VDR/VDRE-driven transcription. In HEK293 kidney and TE85 bone cells, co-transfection of low amounts (1-5ng) of a SIRT1-expression vector elicits a reproducible and statistically significant enhancement (1.3- to 2.6-fold) in transcription mediated by VDREs from the CYP3A4 and cyp24a1 genes, where the magnitude of response to 1,25D-ligand is 6- to 30-fold. Inhibition of SIRT1 via EX-527, or utilization of a SIRT1 loss-of-function mutant (H363Y), resulted in abrogation of SIRT1-mediated VDR potentiation. Studies with a novel, non-acetylatable VDR mutant (K413R) showed that the mutant VDR possesses enhanced responsiveness to 1,25D, in conjunction with reduced, but still significant, sensitivity to exogenous SIRT1, indicating that acetylation of lysine 413 is relevant, but that other acetylated residues in VDR contribute to modulation of its activity. We conclude that the acetylation of VDR comprises a negative feedback loop that attenuates 1,25D-VDR signaling. This regulatory loop is reversed by SIRT1-catalyzed deacetylation of VDR to amplify VDR signaling and 1,25D actions.

Original languageEnglish (US)
JournalJournal of Steroid Biochemistry and Molecular Biology
DOIs
StateAccepted/In press - 2017

Fingerprint

Sirtuin 1
Calcitriol Receptors
Calcitriol
Acetylation
Transcription
Vitamins
Ligands
Retinoid X Receptors
Kidney
Cytochrome P-450 CYP3A
HEK293 Cells

Keywords

  • Acetylation
  • Resveratrol
  • Sirtuin-1
  • Transactivation
  • VDR
  • Vitamin D

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Medicine(all)
  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Endocrinology
  • Clinical Biochemistry
  • Cell Biology

Cite this

SIRT1 enzymatically potentiates 1,25-dihydroxyvitamin D3 signaling via vitamin D receptor deacetylation. / Sabir, Marya S.; Khan, Zainab; Hu, Chengcheng; Galligan, Michael A.; Dussik, Christopher M.; Mallick, Sanchita; Stone, Angelika Dampf; Batie, Shane F.; Jacobs, Elizabeth T.; Whitfield, G. K.; Haussler, Mark R.; Heck, Michael C.; Jurutka, Peter.

In: Journal of Steroid Biochemistry and Molecular Biology, 2017.

Research output: Contribution to journalArticle

Sabir, MS, Khan, Z, Hu, C, Galligan, MA, Dussik, CM, Mallick, S, Stone, AD, Batie, SF, Jacobs, ET, Whitfield, GK, Haussler, MR, Heck, MC & Jurutka, P 2017, 'SIRT1 enzymatically potentiates 1,25-dihydroxyvitamin D3 signaling via vitamin D receptor deacetylation', Journal of Steroid Biochemistry and Molecular Biology. https://doi.org/10.1016/j.jsbmb.2017.06.010
Sabir, Marya S. ; Khan, Zainab ; Hu, Chengcheng ; Galligan, Michael A. ; Dussik, Christopher M. ; Mallick, Sanchita ; Stone, Angelika Dampf ; Batie, Shane F. ; Jacobs, Elizabeth T. ; Whitfield, G. K. ; Haussler, Mark R. ; Heck, Michael C. ; Jurutka, Peter. / SIRT1 enzymatically potentiates 1,25-dihydroxyvitamin D3 signaling via vitamin D receptor deacetylation. In: Journal of Steroid Biochemistry and Molecular Biology. 2017.
@article{fd975092a73448ffbf37c744bf2cbc2e,
title = "SIRT1 enzymatically potentiates 1,25-dihydroxyvitamin D3 signaling via vitamin D receptor deacetylation",
abstract = "The hormonal metabolite of vitamin D, 1,25-dihydroxyvitamin D3 (1,25D), binds to the vitamin D receptor (VDR) and promotes heterodimerization of VDR with a retinoid-X-receptor (RXR) to genomically regulate diverse cellular processes. Herein, it is revealed for the first time that VDR is post-translationally acetylated, and that VDR immunoprecipitated from human embryonic kidney (HEK293) cells displays a dramatic decrease in acetylated receptor in the presence of 1,25D-ligand, sirtuin-1 (SIRT1) deacetylase, or the resveratrol activator of SIRT1. To elucidate the functional significance of VDR deacetylation, vitamin-d-responsive-element (VDRE)-based transcriptional assays were performed to determine if deacetylase overexpression affects VDR/VDRE-driven transcription. In HEK293 kidney and TE85 bone cells, co-transfection of low amounts (1-5ng) of a SIRT1-expression vector elicits a reproducible and statistically significant enhancement (1.3- to 2.6-fold) in transcription mediated by VDREs from the CYP3A4 and cyp24a1 genes, where the magnitude of response to 1,25D-ligand is 6- to 30-fold. Inhibition of SIRT1 via EX-527, or utilization of a SIRT1 loss-of-function mutant (H363Y), resulted in abrogation of SIRT1-mediated VDR potentiation. Studies with a novel, non-acetylatable VDR mutant (K413R) showed that the mutant VDR possesses enhanced responsiveness to 1,25D, in conjunction with reduced, but still significant, sensitivity to exogenous SIRT1, indicating that acetylation of lysine 413 is relevant, but that other acetylated residues in VDR contribute to modulation of its activity. We conclude that the acetylation of VDR comprises a negative feedback loop that attenuates 1,25D-VDR signaling. This regulatory loop is reversed by SIRT1-catalyzed deacetylation of VDR to amplify VDR signaling and 1,25D actions.",
keywords = "Acetylation, Resveratrol, Sirtuin-1, Transactivation, VDR, Vitamin D",
author = "Sabir, {Marya S.} and Zainab Khan and Chengcheng Hu and Galligan, {Michael A.} and Dussik, {Christopher M.} and Sanchita Mallick and Stone, {Angelika Dampf} and Batie, {Shane F.} and Jacobs, {Elizabeth T.} and Whitfield, {G. K.} and Haussler, {Mark R.} and Heck, {Michael C.} and Peter Jurutka",
year = "2017",
doi = "10.1016/j.jsbmb.2017.06.010",
language = "English (US)",
journal = "Journal of Steroid Biochemistry and Molecular Biology",
issn = "0960-0760",
publisher = "Elsevier Limited",

}

TY - JOUR

T1 - SIRT1 enzymatically potentiates 1,25-dihydroxyvitamin D3 signaling via vitamin D receptor deacetylation

AU - Sabir, Marya S.

AU - Khan, Zainab

AU - Hu, Chengcheng

AU - Galligan, Michael A.

AU - Dussik, Christopher M.

AU - Mallick, Sanchita

AU - Stone, Angelika Dampf

AU - Batie, Shane F.

AU - Jacobs, Elizabeth T.

AU - Whitfield, G. K.

AU - Haussler, Mark R.

AU - Heck, Michael C.

AU - Jurutka, Peter

PY - 2017

Y1 - 2017

N2 - The hormonal metabolite of vitamin D, 1,25-dihydroxyvitamin D3 (1,25D), binds to the vitamin D receptor (VDR) and promotes heterodimerization of VDR with a retinoid-X-receptor (RXR) to genomically regulate diverse cellular processes. Herein, it is revealed for the first time that VDR is post-translationally acetylated, and that VDR immunoprecipitated from human embryonic kidney (HEK293) cells displays a dramatic decrease in acetylated receptor in the presence of 1,25D-ligand, sirtuin-1 (SIRT1) deacetylase, or the resveratrol activator of SIRT1. To elucidate the functional significance of VDR deacetylation, vitamin-d-responsive-element (VDRE)-based transcriptional assays were performed to determine if deacetylase overexpression affects VDR/VDRE-driven transcription. In HEK293 kidney and TE85 bone cells, co-transfection of low amounts (1-5ng) of a SIRT1-expression vector elicits a reproducible and statistically significant enhancement (1.3- to 2.6-fold) in transcription mediated by VDREs from the CYP3A4 and cyp24a1 genes, where the magnitude of response to 1,25D-ligand is 6- to 30-fold. Inhibition of SIRT1 via EX-527, or utilization of a SIRT1 loss-of-function mutant (H363Y), resulted in abrogation of SIRT1-mediated VDR potentiation. Studies with a novel, non-acetylatable VDR mutant (K413R) showed that the mutant VDR possesses enhanced responsiveness to 1,25D, in conjunction with reduced, but still significant, sensitivity to exogenous SIRT1, indicating that acetylation of lysine 413 is relevant, but that other acetylated residues in VDR contribute to modulation of its activity. We conclude that the acetylation of VDR comprises a negative feedback loop that attenuates 1,25D-VDR signaling. This regulatory loop is reversed by SIRT1-catalyzed deacetylation of VDR to amplify VDR signaling and 1,25D actions.

AB - The hormonal metabolite of vitamin D, 1,25-dihydroxyvitamin D3 (1,25D), binds to the vitamin D receptor (VDR) and promotes heterodimerization of VDR with a retinoid-X-receptor (RXR) to genomically regulate diverse cellular processes. Herein, it is revealed for the first time that VDR is post-translationally acetylated, and that VDR immunoprecipitated from human embryonic kidney (HEK293) cells displays a dramatic decrease in acetylated receptor in the presence of 1,25D-ligand, sirtuin-1 (SIRT1) deacetylase, or the resveratrol activator of SIRT1. To elucidate the functional significance of VDR deacetylation, vitamin-d-responsive-element (VDRE)-based transcriptional assays were performed to determine if deacetylase overexpression affects VDR/VDRE-driven transcription. In HEK293 kidney and TE85 bone cells, co-transfection of low amounts (1-5ng) of a SIRT1-expression vector elicits a reproducible and statistically significant enhancement (1.3- to 2.6-fold) in transcription mediated by VDREs from the CYP3A4 and cyp24a1 genes, where the magnitude of response to 1,25D-ligand is 6- to 30-fold. Inhibition of SIRT1 via EX-527, or utilization of a SIRT1 loss-of-function mutant (H363Y), resulted in abrogation of SIRT1-mediated VDR potentiation. Studies with a novel, non-acetylatable VDR mutant (K413R) showed that the mutant VDR possesses enhanced responsiveness to 1,25D, in conjunction with reduced, but still significant, sensitivity to exogenous SIRT1, indicating that acetylation of lysine 413 is relevant, but that other acetylated residues in VDR contribute to modulation of its activity. We conclude that the acetylation of VDR comprises a negative feedback loop that attenuates 1,25D-VDR signaling. This regulatory loop is reversed by SIRT1-catalyzed deacetylation of VDR to amplify VDR signaling and 1,25D actions.

KW - Acetylation

KW - Resveratrol

KW - Sirtuin-1

KW - Transactivation

KW - VDR

KW - Vitamin D

UR - http://www.scopus.com/inward/record.url?scp=85021307219&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85021307219&partnerID=8YFLogxK

U2 - 10.1016/j.jsbmb.2017.06.010

DO - 10.1016/j.jsbmb.2017.06.010

M3 - Article

C2 - 28636886

AN - SCOPUS:85021307219

JO - Journal of Steroid Biochemistry and Molecular Biology

JF - Journal of Steroid Biochemistry and Molecular Biology

SN - 0960-0760

ER -