title: Crystallization of the Ets1-Runx1-CBFβ-DNA complex formed on the TCRα gene enhancer.
abstract: Gene transcription is regulated in part through the assembly of multiple transcription factors (TFs) on gene enhancers. To enable examination of the mechanism underlying the formation of these complexes and their response to a phosphorylation signal, two kinds of higher-order TF-DNA assemblies were crystallized composed of an unmodified or phosphorylated Ets1 fragment, a Runx1(L94K) fragment and a CBFβ fragment on the T-cell receptor (TCR) α gene enhancer. Within these complexes, the Ets1 and Runx1 fragments contain intrinsically disordered regulatory regions as well as their DNA-binding domains. Crystals of the complex containing unmodified Ets1 belonged to space group P212121, with unit-cell parameters a = 78.7, b = 102.1, c = 195.0 Å, and diffracted X-rays to a resolution of 2.35 Å, and those containing phosphorylated Ets1 belonged to the same space group, with unit-cell parameters a = 78.6, b = 101.7, c = 194.7 Å, and diffracted X-rays to a similar resolution. To facilitate crystallization, a Runx1 residue involved in a hydrophobic patch that was predicted to be engaged in crystal packing based on the previously reported structures of Runx1-containing crystals was mutated.
authors.affiliation: Department of Biochemistry, Yokohama City University Graduate School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Shiina
authors.affiliation: Department of Biochemistry, Yokohama City University Graduate School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Hamada
authors.affiliation: Department of Biochemistry, Yokohama City University School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan.
authors.forename: Taiko
authors.initials: T
authors.lastname: Inoue-Bungo
authors.affiliation: Department of Biochemistry, Yokohama City University School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan.
authors.forename: Mariko
authors.initials: M
authors.lastname: Shimamura
authors.affiliation: Department of Biochemistry, Yokohama City University School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan.
authors.forename: Shiho
authors.initials: S
authors.lastname: Baba
authors.affiliation: Department of Biochemistry, Yokohama City University Graduate School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan.
authors.forename: Ko
authors.initials: K
authors.lastname: Sato
authors.affiliation: Department of Biochemistry, Yokohama City University Graduate School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Ogata
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D050676:Core Binding Factor Alpha 2 Subunit / Q000737:chemistry*; D003460:Crystallization; D004247:DNA / Q000737:chemistry; D004742:Enhancer Elements, Genetic; D019673:Genes, T-Cell Receptor alpha; D006801:Humans; D046911:Macromolecular Substances / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D050784:Proto-Oncogene Protein c-ets-1 / Q000737:chemistry*
chemical_list: D050676:Core Binding Factor Alpha 2 Subunit; C494041:ETS1 protein, human; D046911:Macromolecular Substances; D050784:Proto-Oncogene Protein c-ets-1; C493726:Runx1 protein, mouse; D004247:DNA
keywords: CBFβ; Ets1; Runx1; TCR gene enhancer; chemical modification; transcription