PMID- 27852823
OWN - NLM
STAT- MEDLINE
DCOM- 20170605
LR  - 20220317
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 292
IP  - 4
DP  - 2017 Jan 27
TI  - Phosphorylation of Human Retinoid X Receptor alpha at Serine 260 Impairs Its 
      Subcellular Localization, Receptor Interaction, Nuclear Mobility, and 
      1alpha,25-Dihydroxyvitamin D3-dependent DNA Binding in Ras-transformed Keratinocytes.
PG  - 1490-1509
LID - 10.1074/jbc.M116.758185 [doi]
AB  - Human retinoid X receptor alpha (hRXRalpha) plays a critical role in DNA binding and 
      transcriptional activity through heterodimeric association with several members 
      of the nuclear receptor superfamily, including the human vitamin D receptor 
      (hVDR). We previously showed that hRXRalpha phosphorylation at serine 260 through the 
      Ras-Raf-MAPK ERK1/2 activation is responsible for resistance to the growth 
      inhibitory effects of 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)), the 
      biologically active metabolite of vitamin D(3) To further investigate the 
      mechanism of this resistance, we studied intranuclear dynamics of hVDR and 
      hRXRalpha-tagged constructs in living cells together with endogenous and tagged 
      protein in fixed cells. We find that hVDR-, hRXRalpha-, and hVDR-hRXRalpha complex 
      accumulate in the nucleus in 1alpha,25(OH)(2)D(3)-treated HPK1A cells but to a lesser 
      extent in HPK1ARas-treated cells. Also, by using fluorescence resonance energy 
      transfer (FRET), we demonstrate increased interaction of the hVDR-hRXRalpha complex 
      in 1alpha,25(OH)(2)D(3)-treated HPK1A but not HPK1ARas cells. In HPK1ARas cells, 
      1alpha,25(OH)(2)D(3)-induced nuclear localization and interaction of hRXRalpha are 
      restored when cells are treated with the MEK1/2 inhibitor UO126 or following 
      transfection of the non-phosphorylatable hRXRalpha Ala-260 mutant. Finally, we 
      demonstrate using fluorescence loss in photobleaching and quantitative 
      co-localization with chromatin that RXR immobilization and co-localization with 
      chromatin are significantly increased in 1alpha,25(OH)(2)D(3)-treated HPK1ARas cells 
      transfected with the non-phosphorylatable hRXRalpha Ala-260 mutant. This suggests 
      that hRXRalpha phosphorylation significantly disrupts its nuclear localization, 
      interaction with VDR, intra-nuclear trafficking, and binding to chromatin of the 
      hVDR-hRXR complex.
CI  - (c) 2017 by The American Society for Biochemistry and Molecular Biology, Inc.
FAU - Jusu, Sylvester
AU  - Jusu S
AD  - From the Department of Medicine, Calcium Research Laboratory, Royal Victoria 
      Hospital, McGill University, Montreal, Quebec H4A 3J1.
AD  - the Department of Medicine, Experimental Therapeutics and Metabolism Program, 
      McGill University Health Center, Montreal, Quebec H4A 3J1, Canada.
FAU - Presley, John F
AU  - Presley JF
AD  - the Department of Anatomy and Cell Biology, McGill University, Montreal, Quebec 
      H3A 0C7, and.
FAU - Kremer, Richard
AU  - Kremer R
AD  - From the Department of Medicine, Calcium Research Laboratory, Royal Victoria 
      Hospital, McGill University, Montreal, Quebec H4A 3J1, richard.kremer@mcgill.ca.
AD  - the Department of Medicine, Experimental Therapeutics and Metabolism Program, 
      McGill University Health Center, Montreal, Quebec H4A 3J1, Canada.
LA  - eng
GR  - MOP10839/CIHR/Canada
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20161116
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Retinoid X Receptor alpha)
RN  - 452VLY9402 (Serine)
RN  - EC 2.7.11.24 (Extracellular Signal-Regulated MAP Kinases)
RN  - EC 3.6.5.2 (ras Proteins)
RN  - FXC9231JVH (Calcitriol)
SB  - IM
MH  - Active Transport, Cell Nucleus/drug effects
MH  - Amino Acid Substitution
MH  - Calcitriol/*pharmacology
MH  - Cell Line, Transformed
MH  - Cell Nucleus/genetics/*metabolism
MH  - Extracellular Signal-Regulated MAP Kinases/genetics/metabolism
MH  - Humans
MH  - Keratinocytes/*metabolism
MH  - Mutation, Missense
MH  - Phosphorylation/drug effects/genetics
MH  - Retinoid X Receptor alpha/genetics/*metabolism
MH  - Serine
MH  - ras Proteins/genetics/metabolism
PMC - PMC5270490
OTO - NOTNLM
OT  - fluorescence loss in photobleaching (FLIP)
OT  - fluorescence resonance energy transfer (FRET)
OT  - mitogen-activated protein kinase (MAPK)
OT  - phosphorylation
OT  - retinoid
OT  - retinoid X receptor
OT  - vitamin D
OT  - vitamin D receptor
EDAT- 2016/11/18 06:00
MHDA- 2017/06/06 06:00
PMCR- 2018/01/27
CRDT- 2016/11/18 06:00
PHST- 2016/09/12 00:00 [received]
PHST- 2016/10/14 00:00 [revised]
PHST- 2016/11/18 06:00 [pubmed]
PHST- 2017/06/06 06:00 [medline]
PHST- 2016/11/18 06:00 [entrez]
PHST- 2018/01/27 00:00 [pmc-release]
AID - S0021-9258(20)32131-1 [pii]
AID - M116.758185 [pii]
AID - 10.1074/jbc.M116.758185 [doi]
PST - ppublish
SO  - J Biol Chem. 2017 Jan 27;292(4):1490-1509. doi: 10.1074/jbc.M116.758185. Epub 
      2016 Nov 16.