PMID- 9169418
OWN - NLM
STAT- MEDLINE
DCOM- 19970626
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 272
IP  - 23
DP  - 1997 Jun 6
TI  - Mutations in the 1,25-dihydroxyvitamin D3 receptor identifying C-terminal amino 
      acids required for transcriptional activation that are functionally dissociated 
      from hormone binding, heterodimeric DNA binding, and interaction with basal 
      transcription factor IIB, in vitro.
PG  - 14592-9
AB  - To investigate a potential ligand-dependent transcriptional activation domain 
      (AF-2) in the C-terminal region of the human vitamin D receptor (hVDR), two 
      conserved residues, Leu-417 and Glu-420, were replaced with alanines by 
      site-directed mutagenesis (L417A and E420A). Transcriptional activation in 
      response to 1, 25-dihydroxyvitamin D3 (1,25-(OH)2D3) was virtually eliminated 
      when either point mutant was transfected into several mammalian cell lines. 
      Furthermore, both mutants exhibited a dominant negative phenotype when expressed 
      in COS-7 cells. Scatchard analysis at 4 degrees C and a ligand-dependent DNA 
      binding assay at 25 degrees C revealed essentially normal 1,25-(OH)2D3 binding 
      for the mutant hVDRs, which were also equivalent to native receptor in 
      associating with the rat osteocalcin vitamin D responsive element as a presumed 
      heterodimer with retinoid X receptor. Glutathione S-transferase-human 
      transcription factor IIB (TFIIB) fusion protein linked to Sepharose equally 
      coprecipitated the wild-type hVDR and the AF-2 mutants. These data implicate 
      amino acids Leu-417 and Glu-420, residing in a putative alpha-helical region at 
      the extreme C terminus of hVDR, as critical in the mechanism of 1, 
      25-(OH)2D3-stimulated transcription, likely mediating an interaction with a 
      coactivator(s) or a component of the basal transcriptional machinery distinct 
      from TFIIB.
FAU - Jurutka, P W
AU  - Jurutka PW
AD  - Department of Biochemistry, College of Medicine, The University of Arizona, 
      Tucson, Arizona 85724, USA.
FAU - Hsieh, J C
AU  - Hsieh JC
FAU - Remus, L S
AU  - Remus LS
FAU - Whitfield, G K
AU  - Whitfield GK
FAU - Thompson, P D
AU  - Thompson PD
FAU - Haussler, C A
AU  - Haussler CA
FAU - Blanco, J C
AU  - Blanco JC
FAU - Ozato, K
AU  - Ozato K
FAU - Haussler, M R
AU  - Haussler MR
LA  - eng
GR  - AR-15781/AR/NIAMS NIH HHS/United States
GR  - DK-33351/DK/NIDDK NIH HHS/United States
GR  - DK-49604/DK/NIDDK NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Receptors, Calcitriol)
RN  - 0 (Receptors, Retinoic Acid)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Retinoid X Receptors)
RN  - 0 (Transcription Factor TFIIB)
RN  - 0 (Transcription Factors)
RN  - 104982-03-8 (Osteocalcin)
RN  - 3KX376GY7L (Glutamic Acid)
RN  - 9007-49-2 (DNA)
RN  - FXC9231JVH (Calcitriol)
RN  - GMW67QNF9C (Leucine)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Binding Sites
MH  - COS Cells
MH  - Calcitriol/pharmacology
MH  - Conserved Sequence
MH  - DNA/*metabolism
MH  - Dimerization
MH  - Glutamic Acid
MH  - Humans
MH  - Kinetics
MH  - Leucine
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Osteocalcin/biosynthesis
MH  - Phenotype
MH  - Point Mutation
MH  - Protein Structure, Secondary
MH  - Rats
MH  - Receptors, Calcitriol/biosynthesis/chemistry/*metabolism
MH  - Receptors, Retinoic Acid/chemistry
MH  - Recombinant Proteins/biosynthesis/chemistry/metabolism
MH  - Retinoid X Receptors
MH  - Sequence Alignment
MH  - Sequence Homology, Amino Acid
MH  - Transcription Factor TFIIB
MH  - Transcription Factors/chemistry/*metabolism
MH  - *Transcriptional Activation
MH  - Transfection
EDAT- 1997/06/06 00:00
MHDA- 1997/06/06 00:01
CRDT- 1997/06/06 00:00
PHST- 1997/06/06 00:00 [pubmed]
PHST- 1997/06/06 00:01 [medline]
PHST- 1997/06/06 00:00 [entrez]
AID - S0021-9258(19)62433-6 [pii]
AID - 10.1074/jbc.272.23.14592 [doi]
PST - ppublish
SO  - J Biol Chem. 1997 Jun 6;272(23):14592-9. doi: 10.1074/jbc.272.23.14592.