DNA binding properties of the vitamin D3 receptor zinc finger region.
The DNA binding domains of the nuclear receptor superfamily are highly conserved and consist of residues that fold into two zinc finger-like motifs, suggesting that the structures of this region among the members of the superfamily are likely to be very similar. Furthermore, the response elements that these receptors bind to are similar in sequence and organization. Nevertheless, these receptors selectively recognize target response elements and differentially regulate linked genes. In order to study the details of receptor:DNA binding, we have overexpressed and purified the vitamin D3 receptor DNA binding domain (VDRF) and have begun characterizing its DNA binding properties. We find that the VDRF protein binds strongly and specifically to direct repeats constituting a vitamin D response element from the mouse osteopontin (Spp-1) promoter region but weakly to the human osteocalcin vitamin D response element. Unlike receptors that recognize hormone response elements oriented as inverted repeats, such as the glucocorticoid receptor (GR) and estrogen receptor, VDRF appears to bind half-sites noncooperatively, without the free energy contribution of dimerization seen when the glucocorticoid receptor DNA binding domain associates with a glucocorticoid response element. By comparing and contrasting the DNA binding properties of the vitamin D and glucocorticoid receptors, we suggest a model for how receptors that prefer direct repeats differ in their binding strategy from those that recognize inverted repeats.
Freedman
L P
LP
Cell Biology and Genetics Program, Sloan-Kettering Institute, New York, New York 10021.
Towers
T L
TL
D000595:Amino Acid Sequence; D001483:Base Sequence; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D004926:Escherichia coli; D015870:Gene Expression / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D053495:Osteopontin; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D018167:Receptors, Calcitriol; D011965:Receptors, Glucocorticoid / Q000235:genetics; D011987:Receptors, Steroid / Q000235:genetics / Q000502:physiology*; D012795:Sialoglycoproteins / Q000235:genetics; D016335:Zinc Fingers / Q000235:genetics / Q000502:physiology*
D004268:DNA-Binding Proteins; D018167:Receptors, Calcitriol; D011965:Receptors, Glucocorticoid; D011987:Receptors, Steroid; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; D053495:Osteopontin; D004247:DNA