PMID- 9010343
OWN - NLM
STAT- MEDLINE
DCOM- 19970225
LR  - 20190831
IS  - 0960-0760 (Print)
IS  - 0960-0760 (Linking)
VI  - 59
IP  - 5-6
DP  - 1996 Dec
TI  - Vitamin D receptor interactions with the murine osteopontin response element.
PG  - 377-88
AB  - The nature of the DNA binding interactions of the human vitamin D receptor (hVDR) 
      with the murine osteopontin vitamin D response element (mOP VDRE) was examined. 
      Both recombinant hVDR and human retinoid X receptor beta (hRXRbeta) proteins were 
      obtained from baculovirus-infected Sf9 insect cells. Mixing extracts of the two 
      recombinant proteins resulted in the strong, specific formation of a slower 
      migrating complex in the electrophoretic mobility shift assay. Crude extracts of 
      the expressed hVDR alone were also capable of binding with high affinity to the 
      mOP sequence, and this binding was enhanced in the presence of 
      1,25-dihydroxyvitamin D3 (1,25-(OH)2D3). Competition experiments confirmed the 
      specificity of this interaction and revealed that the human osteocalcin VDRE was 
      a poor competitor for this binding. Ethylation interference footprint analyses of 
      hVDR/hRXRbeta and hVDR complexes revealed only subtle differences in how these 
      two different VDR-containing complexes interacted with the mOP VDRE. The 
      footprints displayed contact points in both halves of the direct repeat format, 
      confirming the dimeric and major groove interactions of both types of complexes. 
      DNA affinity chromatography of labelled hVDR extracts revealed a peak eluting at 
      ca. 290 mM KC1 that was capable of rebinding to the mOP sequence in gel shift 
      experiments. Ultraviolet (UV) light-crosslinking experiments of hVDR extracts 
      alone to radiolabelled DNA were consistent with the existence of a homodimeric 
      hVDR interaction. Additionally, these experiments confirmed the direct 
      interaction of a hVDR/hRXRbeta heterodimer when mixed extracts were utilized. 
      From these results we infer that homodimers of the hVDR which respond with 
      enhanced DNA binding to particular vitamin D response elements when exposed to 
      1,25-(OH)2D3 are possible. This may be of functional significance when RXR 
      proteins are limiting or RXR ligand is present within a cell.
FAU - Koszewski, N J
AU  - Koszewski NJ
AD  - University of Kentucky Medical Center, Department of Internal Medicine, Lexington 
      40536-0084, USA.
FAU - Reinhardt, T A
AU  - Reinhardt TA
FAU - Horst, R L
AU  - Horst RL
LA  - eng
GR  - DK47883/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - J Steroid Biochem Mol Biol
JT  - The Journal of steroid biochemistry and molecular biology
JID - 9015483
RN  - 0 (Cross-Linking Reagents)
RN  - 0 (Receptors, Calcitriol)
RN  - 0 (Receptors, Retinoic Acid)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Retinoid X Receptors)
RN  - 0 (SPP1 protein, human)
RN  - 0 (Sialoglycoproteins)
RN  - 0 (Spp1 protein, mouse)
RN  - 0 (Transcription Factors)
RN  - 106441-73-0 (Osteopontin)
RN  - FXC9231JVH (Calcitriol)
SB  - IM
MH  - Animals
MH  - Baculoviridae/genetics
MH  - Binding Sites
MH  - Binding, Competitive
MH  - Blotting, Western
MH  - Calcitriol/metabolism
MH  - Cross-Linking Reagents
MH  - Cytosol/metabolism
MH  - Electrophoresis
MH  - Humans
MH  - Insecta/genetics/metabolism/virology
MH  - Mice
MH  - Osteopontin
MH  - Protein Conformation
MH  - Receptors, Calcitriol/*chemistry/genetics/*metabolism
MH  - Receptors, Retinoic Acid/immunology/metabolism
MH  - Recombinant Proteins/chemistry/genetics/metabolism
MH  - Retinoid X Receptors
MH  - Sialoglycoproteins/chemistry/genetics/*metabolism
MH  - Time Factors
MH  - Transcription Factors/immunology/metabolism
MH  - Ultraviolet Rays
EDAT- 1996/12/01 00:00
MHDA- 1996/12/01 00:01
CRDT- 1996/12/01 00:00
PHST- 1996/12/01 00:00 [pubmed]
PHST- 1996/12/01 00:01 [medline]
PHST- 1996/12/01 00:00 [entrez]
AID - 10.1016/s0960-0760(96)00127-6 [doi]
PST - ppublish
SO  - J Steroid Biochem Mol Biol. 1996 Dec;59(5-6):377-88. doi: 
      10.1016/s0960-0760(96)00127-6.