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Crystallization and preliminary X‐ray analysis of a complex of the FOXO1 and Ets1 DNA‐binding domains and DNA
Author(s) -
Choy Wing W.,
Datta Drishadwatti,
Geiger Catherine A.,
Birrane Gabriel,
Grant Marianne A.
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x13024795
Subject(s) - crystallization , dna , ets1 , chemistry , computational biology , microbiology and biotechnology , biophysics , physics , biology , genetics , transcription factor , gene , thermodynamics
The Ets1 transcription factor is a member of the Ets protein family, a group of evolutionarily related DNA‐binding transcriptional factors. Ets proteins activate or repress the expression of genes that are involved in various biological processes, including cellular proliferation, differentiation, development, transformation and apoptosis. FOXO1 is a member of the forkhead‐box proteins (FOX proteins), which comprise a large family of functionally diverse transcription factors involved in cellular proliferation, transformation and differentiation. The FOXO subgroup of FOX proteins regulates the transcription of genes that control metabolism, cell survival, cellular proliferation, DNA damage responses, stress resistance and longevity. The DNA‐binding domains (DBDs) of Ets1 and FOXO1 were crystallized in complex with DNA containing a composite sequence for a noncanonical forkhead binding site (AATAACA) and an ETS site (GGAA), FOX:ETS, by the sitting‐drop vapor‐diffusion method. The FOX:ETS motif has been shown to be a conserved cis ‐acting element in several endothelial cell‐specific genes, including Vegfr2, Tie2, Mef2c and ve‐cadherin. Crystals were grown at 291 K using 30% polyethylene glycol 400, 50 m M Tris pH 8.5, 100 m M KCl, 10 m M MgCl 2 as the reservoir solution. The crystals belonged to space group C 222 1 , with unit‐cell parameters a = 68.7, b = 104.9, c = 136.3 Å. Diffraction data were collected to a resolution of 2.2 Å.