PMID- 9705070
OWN - NLM
STAT- MEDLINE
DCOM- 19981015
LR  - 20190813
IS  - 0303-7207 (Print)
IS  - 0303-7207 (Linking)
VI  - 139
IP  - 1-2
DP  - 1998 Apr 30
TI  - Characterization of the activation function-2 domain of the human 
      1,25-dihydroxyvitamin D3 receptor.
PG  - 15-24
AB  - In this study, we determined the ligand-dependent activation function domain 2 
      (AF-2) of the human vitamin D receptor (hVDR) and characterized it using 
      site-directed mutagenesis. A single mutation at glutamic acid-420 (E420Q) and an 
      additional mutation at leucine-417 (L417A-E420Q) eliminated ligand-dependent 
      transcriptional activation. In addition, lysine-264 was also demonstrated to be 
      vital for ligand-induced transactivation. However, bacterial-overexpressed 
      transcriptional factor IIB (TFIIB) was able to bind to both AF-2 and lysine-264 
      mutant hVDRs in vitro. The ligand-dependent transactivation via wild type hVDR 
      was interfered with weakly only when a 10-fold molar excess of L417A-E420Q 
      plasmid was co-transfected. This suppressive effect was diminished by introducing 
      an additional mutation at a cysteine residue in the DNA binding domain. Thus, we 
      conclude that the AF-2 domain of the hVDR located between amino acids 417 and 
      420, as well as lysine-264, are essential for ligand-dependent transactivation, 
      and that TFIIB was not necessary for the function of these two regions of the 
      hVDR. Our finding that AF-2 mutant hVDRs exhibit only very weak suppressive 
      effect may indicate a difference in the molecular mechanism of the VDR-mediated 
      transactivation from other nuclear receptors.
FAU - Nakajima, S
AU  - Nakajima S
AD  - Department of Environmental Medicine, Research Institute, Osaka Medical Center 
      for Maternal and Child Health, Izumi, Japan.
FAU - Yamagata, M
AU  - Yamagata M
FAU - Sakai, N
AU  - Sakai N
FAU - Ozono, K
AU  - Ozono K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Ireland
TA  - Mol Cell Endocrinol
JT  - Molecular and cellular endocrinology
JID - 7500844
RN  - 0 (Ligands)
RN  - 0 (Receptors, Calcitriol)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Transcription Factor TFIIB)
RN  - 0 (Transcription Factors)
RN  - 104982-03-8 (Osteocalcin)
RN  - 9007-49-2 (DNA)
RN  - K3Z4F929H6 (Lysine)
SB  - IM
MH  - Animals
MH  - COS Cells
MH  - DNA/metabolism
MH  - Humans
MH  - Ligands
MH  - Lysine/physiology
MH  - Osteocalcin/genetics
MH  - Osteosarcoma
MH  - Point Mutation
MH  - Rats
MH  - Receptors, Calcitriol/*genetics/metabolism
MH  - Recombinant Fusion Proteins
MH  - Sequence Alignment
MH  - Transcription Factor TFIIB
MH  - Transcription Factors/metabolism
MH  - Transcriptional Activation/*genetics
MH  - Transfection
MH  - Tumor Cells, Cultured
EDAT- 1998/08/15 00:00
MHDA- 1998/08/15 00:01
CRDT- 1998/08/15 00:00
PHST- 1998/08/15 00:00 [pubmed]
PHST- 1998/08/15 00:01 [medline]
PHST- 1998/08/15 00:00 [entrez]
AID - S0303-7207(98)00077-X [pii]
AID - 10.1016/s0303-7207(98)00077-x [doi]
PST - ppublish
SO  - Mol Cell Endocrinol. 1998 Apr 30;139(1-2):15-24. doi: 
      10.1016/s0303-7207(98)00077-x.