Crystallization and preliminary X-ray analysis of a domain in the Runx2 transcription factor that interacts with the 1?,25 dihydroxy vitamin D3 receptor

Bruna, C; Arriagada, G; Bunster, M; Martinez-Oyanedel, J; Montecino, M.; Lian J.B.; Stein G.S.

Keywords: diffusion, overexpression, mouse, tertiary, animals, expression, transcription, x-ray, binding, core, protein, crystallization, cell, gene, alpha, mice, ray, experiment, subunit, coli, calcitriol, article, factor, analysis, runx2, vapor, method, controlled, animal, study, 1, priority, nonhuman, journal, Receptors,, X, Escherichia, Densitometry,, Structure,, drop, macrogol, Hanging


The Runx2 transcription factor is a key regulator of osteoblast differentiation. In response to 1?,25 dihydroxy vitamin D3, Runx2 may interact with the 1?,25 dihydroxy vitamin D3 receptor (VDR) in the promoter of target genes, producing a synergic activation of their transcription. Previous studies have suggested that the motifs responsible for the VDR-Runx2 interaction are contained within the 230-361 domain of Runx2. In this work, we confirmed by GST-pull down that Runx2l(209-361) is sufficient to interact with the VDR. To obtain structural information, GST-Runx2l(209-361) protein was overexpressed in Escherichia coli, purified and crystallized using the hanging-drop vapor-diffusion method and polyethyleneglycol as a precipitant. The crystals were found to diffract to a maximum resolution of 2.7 Å and a complete data set to a 3.3 Å resolution was collected and analyzed. The crystals belong to the tetragonal system, with a space group P4 and unit-cell parameters of a = b = 90.8, and c = 57.2 Å. The presence of a monomer of the recombinant GST-Runx2 l(209-361) in the asymmetric unit gives a VM of 2.7 Å3 Da-1 and a solvent content of 54.8%. © 2007 Wiley-Liss, Inc.

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Volumen: 101
Número: 3
Editorial: Wiley
Fecha de publicación: 2007
Página de inicio: 785
Página final: 789