Open AccessPreparation, crystallization and preliminary X‐ray diffraction analysis of the DNA‐binding domain of the Ets transcription factor in complex with target DNA
Author(s) -
Suwa Yoshiaki,
Nakamura Teruya,
Toma Sachiko,
Ikemizu Shinji,
Kai Hirofumi,
Yamagata Yuriko
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108002662
Subject(s) - transcription factor , dna binding domain , dna , microbiology and biotechnology , transcription (linguistics) , cleavage (geology) , crystallization , promoter , chemistry , crystallography , biology , gene , biochemistry , gene expression , linguistics , philosophy , paleontology , organic chemistry , fracture (geology)
The Ets2 transcription factor is a member of the Ets transcription‐factor family. Ets2 plays a role in the malignancy of cancer and in Down's syndrome by regulating the transcription of various genes. The DNA‐binding domain of Ets2 (Ets domain; ETSD), which contains residues that are highly conserved among Ets transcription‐factor family members, was expressed as a GST‐fusion protein. The aggregation of ETSD produced after thrombin cleavage could be prevented by treatment with NDSB‐195 (nondetergent sulfobetaine 195). ETSD was crystallized in complex with DNA containing the Ets2 target sequence (GGAA) by the hanging‐drop vapour‐diffusion method. The best crystals were grown using 25% PEG 3350, 80 m M magnesium acetate, 50 m M sodium cacodylate pH 5.0/5.5 as the reservoir at 293 K. The crystals belonged to space group C 2, with unit‐cell parameters a = 85.89, b = 95.52, c = 71.89 Å, β = 101.7° and a V M value of 3.56 Å 3 Da −1 . Diffraction data were collected to a resolution of 3.0 Å.