Crystallographic analysis of the interaction of the glucocorticoid receptor with DNA

Luisi, Ben F.; Xu, Wei; Otwinowski, Zbyszek; Freedman, Leonard P.; Yamamoto, Keith R.; Sigler, Paul B.

doi:10.1038/352497a0

Cited by 1,445 publications

(1,046 citation statements)

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“…Also, there are fewer direct contacts between the protein and the DNA, and the DNA lacks major protein-induced changes in conformation. In the present case, the protein is much further removed from the DNA than is the case with either the glucocorticoid receptor or EcoRV endonuclease bound to noncognate DNA (Luisi et al, 1991;WinMer et al, 1993;Gewirth & Sigler, 1995). In this respect, the postulated Cro complex is more similar to that proposed for the nonspecific binding of CAP (Weber & Steitz, 1984), except that the protein is arranged asymmetrically with respect to the DNA.…”

Section: Proposed Model For the Binding Of Wild-type Cro To Noncognatmentioning

confidence: 53%

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Crystal structure of an engineered cro monomer bound nonspecifically to DNA: Possible implications for nonspecific binding by the wild‐type protein

1998

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The structure has been determined at 3.0 8, resolution of a complex of engineered monomeric Cro repressor with a seven-base pair DNA fragment. Although the sequence of the DNA corresponds to the consensus half-operator that is recognized by each subunit of the wild-type Cro dimer, the complex that is formed in the crystals by the isolated monomer appears to correspond to a sequence-independent mode of association. The overall orientation of the protein relative to the DNA is markedly different from that observed for Cro dimer bound to a consensus operator. The recognition helix is rotated 48" further out of the major groove, while the turn region of the helix-turn-helix remains in contact with the DNA backbone. All of the direct base-specific interactions seen in the wild-type Cro-operator complex are lost. Virtually all of the ionic interactions with the DNA backbone, however, are maintained, as is the subset of contacts between the DNA backbone and a channel on the protein surface. Overall, 25% less surface area is buried at the protein-DNA interface than for half of the wild-type Cro-operator complex, and the contacts are more ionic in character due to a reduction of hydrogen bonding and van der Waals interactions. Based on this crystal structure, model building was used to develop a possible model for the sequence-nonspecific interaction of the wild-type Cro dimer with DNA. In the sequence-specific complex, the DNA is bent, the protein dimer undergoes a large hinge-bending motion relative to the uncomplexed form, and the complex is twofold symmetric. In contrast, in the proposed nonspecific complex the DNA is straight, the protein retains a conformation similar to the apo form, and the complex lacks twofold symmetry. The model is consistent with thermodynamic, chemical, and mutagenic studies, and suggests that hinge bending of the Cro dimer may be critical in permitting the transition from the binding of protein at generic sites on the DNA to binding at high affinity operator sites.

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Section: Proposed Model For the Binding Of Wild-type Cro To Noncognatmentioning

confidence: 53%

“…To date, there is relatively little information on the structure of such complexes (Luisi et al, 1991;Winkler et al, 1993;Gewirth & Sigler, 1995).…”

mentioning

confidence: 99%

Crystal structure of an engineered cro monomer bound nonspecifically to DNA: Possible implications for nonspecific binding by the wild‐type protein

1998

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“…However, a certain degree of specificity is retained in the N-terminal zinc finger: For instance, three amino acid residues in this zinc finger are essential for the specificity of the DNA binding domain of the GR and the estrogen receptor (Green et al, 1988;Mader et al, 1989;Danielsen et al, 1989;Umesono and Evans, 1989). The N-terminal zinc finger binds to specific nucleotides of the hormone responsive element while the C-terminal zinc finger is more important for stabilization of the protein-DNA complex (Chalepakis et al, 1988;Hard et al, 1990;Luisi et al, 1991). The DNA binding domain of the GR also contains a second nuclear translocation signal.…”

Section: Primary Structurementioning

confidence: 99%

Mineralocorticoid and glucocorticoid receptors in the brain. Implications for ion permeability and transmitter systems

Joëls

Kloet

1994

Progress in Neurobiology

368

168

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“…The DNAbinding domain binds specifically to hormone-responsive elements in DNA. The structural basis of DNA recognition is well established from nuclear magnetic resonance and X-ray crystallography analyses [10][11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

Large-scale expression and purification of the human vitamin D receptor and its ligand-binding domain for structural studies

Juntunen

Rochel

Moras

et al. 1999

Biochemical Journal

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We have expressed recombinant human vitamin D receptor and its ligand-binding domain in Spodoptera frugiperda (Sf9) insect cells with a 30-litre bioreactor. Both proteins were purified to apparent homogeneity with yields of 0.5-3.5 mg\l. Gel-filtration analyses indicated that the purified human vitamin D receptor and its ligand-binding domain were present as monomers in solution. The purified vitamin D receptor and its ligand-binding domain were demonstrated to bind 1α,25-dihydroxyvitamin D $ with high affinity, the K d values ranging from 0.9 to 1.2 nM. Neutron scattering studies of the ligand-binding domain demonstrated that the samples are homogeneous and contain monomeric species of polypeptides. The purified vitamin D receptor

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Crystallographic analysis of the interaction of the glucocorticoid receptor with DNA

Cited by 1,445 publications

References 50 publications

Crystal structure of an engineered cro monomer bound nonspecifically to DNA: Possible implications for nonspecific binding by the wild‐type protein

Crystal structure of an engineered cro monomer bound nonspecifically to DNA: Possible implications for nonspecific binding by the wild‐type protein

Mineralocorticoid and glucocorticoid receptors in the brain. Implications for ion permeability and transmitter systems

Large-scale expression and purification of the human vitamin D receptor and its ligand-binding domain for structural studies

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