Crystallization and preliminary X-ray analysis of a complex of the FOXO1 and Ets1 DNA-binding domains and DNA
Crystallization and preliminary X-ray analysis of a complex of the FOXO1 and Ets1 DNA-binding domains and DNA
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DOI：
10.1107/s2053230x13024795
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发表时间：
2014-01-01
影响因子：
0.9
通讯作者：
Grant, Marianne A.
中科院分区：
文献类型：
--
作者：
Choy, Wing W.;Datta, Drishadwatti;Grant, Marianne A.
Ets1 转录因子是 Ets 蛋白家族的成员，Ets 蛋白家族是一组进化相关的 DNA 结合转录因子。 Ets蛋白激活或抑制参与各种生物过程的基因表达，包括细胞增殖、分化、发育、转化和凋亡。 FOXO1 是叉头盒蛋白（FOX 蛋白）的成员，FOX 蛋白由参与细胞增殖、转化和分化的功能多样的转录因子大家族组成。 FOX 蛋白的 FOXO 亚组调节控制新陈代谢、细胞存活、细胞增殖、DNA 损伤反应、应激抵抗和长寿的基因转录。通过坐滴蒸气扩散法，Ets1 和 FOXO1 的 DNA 结合域 (DBD) 与含有非规范叉头结合位点 (AATAACA) 和 ETS 位点 (GGAA)、FOX: ETS 复合序列的 DNA 形成复合物。 FOX:ETS 基序已被证明是多个内皮细胞特异性基因（包括 Vegfr2、Tie2、Mef2c 和 ve-cadherin）中的保守顺式作用元件。使用 30% 聚乙二醇 400、50 mM Tris pH 8.5、100 mM KCl、10 mM MgCl2 作为储存溶液，在 291 K 下生长晶体。该晶体属于空间群C222(1)，晶胞参数a = 68.7，b = 104.9，c = 136.3 埃。收集分辨率为 2.2 埃的衍射数据。
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 mM Tris pH 8.5, 100 mM KCl, 10 mM MgCl2 as the reservoir solution. The crystals belonged to space group C222(1), with unit-cell parameters a = 68.7, b = 104.9, c = 136.3 angstrom. Diffraction data were collected to a resolution of 2.2 angstrom.