Functional and structural characterization of the insertion region in the ligand binding domain of the vitamin D nuclear receptor.
Vitamin D nuclear receptor mediates the genomic actions of the active form of vitamin D, 1,25(OH)2D3. This hormone is involved in calcium and phosphate metabolism and cell differentiation. Compared to other nuclear receptors, VDR presents a large insertion region at the N-terminal part of the ligand binding domain between helices H1 and H3, encoded by an additional exon. This region is poorly conserved in VDR in different species and is not well ordered as observed by secondary structure prediction. We engineered a VDR ligand binding domain mutant by removing this insertion region. Here we report its biochemical and biophysical characterization. The mutant protein exhibits the same ligand binding, dimerization with retinoid X receptor and transactivation properties as the wild-type VDR, suggesting that the insertion region does not affect these main functions. Solution studies by small angle X-ray scattering shows that the conformation in solution of the VDR mutant is similar to that observed in the crystal and that the insertion region in the VDR wild-type is not well ordered.
Rochel
N
N
Laboratoire de Biologie et Génomique Structurale, UPR 9004, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, Illkirch Cedex, France.
Tocchini-Valentini
G
G
Egea
P F
PF
Juntunen
K
K
Garnier
J M
JM
Vihko
P
P
Moras
D
D
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002117:Calcitriol / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D018167:Receptors, Calcitriol / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D015533:Transcriptional Activation; D014162:Transfection; D014961:X-Ray Diffraction
D008024:Ligands; D018167:Receptors, Calcitriol; D002117:Calcitriol