PMID- 10587460
OWN - NLM
STAT- MEDLINE
DCOM- 20000110
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 38
IP  - 49
DP  - 1999 Dec 7
TI  - Characterization of unique DNA-binding and transcriptional-activation functions 
      in the carboxyl-terminal extension of the zinc finger region in the human vitamin 
      D receptor.
PG  - 16347-58
AB  - The vitamin D receptor (VDR) binds 1,25-dihydroxyvitamin D(3) and mediates its 
      actions on gene transcription by heterodimerizing with retinoid X receptors 
      (RXRs) on direct repeat (DR+3) vitamin D responsive elements (VDREs) located in 
      target genes. The VDRE binding function of VDR has been primarily ascribed to the 
      zinc finger region (residues 24-87). To define the minimal VDRE binding domain 
      for human VDR (hVDR), a series of C-terminally truncated hVDR mutants (Delta134, 
      Delta113, Delta102, Delta90, Delta84, Delta80, and Delta60) was generated and 
      expressed in bacteria. Only the Delta134 and Delta113 mutants bound the VDRE 
      (predominantly as monomers), suggesting that, in addition to the conserved zinc 
      finger region of hVDR, as many as 25 amino acids in a C-terminal extension (CTE) 
      participate in DNA binding. Site-directed mutagenesis of conserved charged 
      residues in full-length hVDR was then performed to dissect the functional 
      significance of the CTE (residues 88-112) in the context of the complete 
      hVDR-RXR-VDRE interaction. Functional assays revealed that E98K/E99K, 
      R102A/K103A/R104A, and K109A/R110A/K111A mutant hVDRs possessed dramatically 
      reduced DNA binding and transcriptional activities, whereas distinct point 
      mutants, such as K103A, bound to DNA normally but lacked transcriptional 
      activity. Therefore, the boundary for the minimal DNA-binding domain in hVDR 
      extends C-terminal of the zinc fingers to Lys-111, with clusters of highly 
      conserved charged amino acids playing a crucial role in binding to the DR+3 
      element. Further, individual residues in this region (e.g., Lys-103) may lie on 
      the opposing face of a DNA-binding alpha-helix, where they could contact 
      transcriptional coactivators or basal transcription factors.
FAU - Hsieh, J C
AU  - Hsieh JC
AD  - Department of Biochemistry, College of Medicine, The University of Arizona, 
      Tucson 85724, USA.
FAU - Whitfield, G K
AU  - Whitfield GK
FAU - Oza, A K
AU  - Oza AK
FAU - Dang, H T
AU  - Dang HT
FAU - Price, J N
AU  - Price JN
FAU - Galligan, M A
AU  - Galligan MA
FAU - Jurutka, P W
AU  - Jurutka PW
FAU - Thompson, P D
AU  - Thompson PD
FAU - Haussler, C A
AU  - Haussler CA
FAU - Haussler, M R
AU  - Haussler MR
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (Receptors, Calcitriol)
RN  - 0 (Trans-Activators)
SB  - IM
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution/genetics
MH  - Conserved Sequence
MH  - DNA-Binding Proteins/*chemistry/genetics/physiology
MH  - Dimerization
MH  - Humans
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Peptide Fragments/*chemistry/genetics/physiology
MH  - Point Mutation
MH  - Protein Structure, Tertiary/genetics
MH  - Receptors, Calcitriol/*chemistry/genetics/physiology
MH  - Sequence Deletion
MH  - Trans-Activators/*chemistry/genetics/physiology
MH  - *Zinc Fingers/genetics
EDAT- 1999/12/10 00:00
MHDA- 1999/12/10 00:01
CRDT- 1999/12/10 00:00
PHST- 1999/12/10 00:00 [pubmed]
PHST- 1999/12/10 00:01 [medline]
PHST- 1999/12/10 00:00 [entrez]
AID - bi9916574 [pii]
AID - 10.1021/bi9916574 [doi]
PST - ppublish
SO  - Biochemistry. 1999 Dec 7;38(49):16347-58. doi: 10.1021/bi9916574.