INTERACTION OF MURINE ETS-1 WITH GGA-BINDING SITES ESTABLISHES THE ETS DOMAIN AS A NEW DNA-BINDING MOTIF
INTERACTION OF MURINE ETS-1 WITH GGA-BINDING SITES ESTABLISHES THE ETS DOMAIN AS A NEW DNA-BINDING MOTIF
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DOI：
10.1101/gad.6.6.975
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发表时间：
1992-06-01
影响因子：
10.5
通讯作者：
GRAVES, BJ
中科院分区：
文献类型：
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作者：
NYE, JA;PETERSEN, JM;GRAVES, BJ
原癌基因ets-1是真核生物转录调节因子新家族的创始成员。使用在大肠杆菌中表达的小鼠ets-1 cDNA的缺失突变体，我们表明，DNA结合结构域对应密切的ETS结构域，一个85个氨基酸的区域，是保守的ets家族成员之间。为了研究ETS结构域的DNA结合的特异性，我们通过化学保护和干扰测定绘制了单体Ets-1片段的DNA接触。DNA骨架相互作用跨越20个核苷酸的区域，并位于螺旋的一面。紧密的磷酸盐和碱基接触限于10个中心核苷酸。联系人地图的主要沟在该网站的中心。侧翼小沟相互作用也被预测。为了确定紧密接触区的序列偏好，我们使用纯化的Ets-1羧基末端片段选择了一组高亲和力结合位点。我们的Ets-1选择的共有序列5 '-A/GCCGGAA/TGT/C-3'不同于果蝇ETS结构域蛋白E74 A的结合共有序列，表明ets家族成员的特异性作用是由ETS结构域介导的。与其他类型的DNA结合蛋白相比，Ets-1产生独特的DNA接触模式。这些研究表明，ETS结构域蛋白结合DNA在一个新的
The proto-oncogene ets-1 is the founding member of a new family of eukaryotic transcriptional regulators. Using deletion mutants of murine ets-1 cDNA expressed in Escherichia coli, we show that the DNA-binding domain corresponds closely to the ETS domain, an 85-amino-acid region that is conserved among ets family members. To investigate the specificity of DNA binding of the ETS domain, we mapped the DNA contacts of a monomeric Ets-1 fragment by chemical protection and interference assays. DNA backbone interactions span a 20-nucleotide region and are localized on one face of the helix. Close phosphate and base contacts are restricted to 10 central nucleotides. Contacts map to the major groove in the center of the site. Flanking minor groove interactions also are predicted. To determine the sequence preference in the close contact zone, we selected a pool of high-affinity binding sites using a purified Ets-1 carboxy-terminal fragment. Our Ets-1-selected consensus, 5'-A/GCCGGAA/TGT/C-3', differs from the binding consensus for the Drosophila ETS domain protein E74A, suggesting that specificity of action of ets family members is mediated by the ETS domain. Compared to other well-characterized classes of DNA-binding proteins, Ets-1 produces a unique pattern of DNA contacts. These studies demonstrate that the ETS domain proteins bind DNA in a novel