DNA-BINDING SPECIFICITY OF GATA FAMILY TRANSCRIPTION FACTORS
DNA-BINDING SPECIFICITY OF GATA FAMILY TRANSCRIPTION FACTORS
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DOI：
10.1128/mcb.13.7.3999
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发表时间：
1993-07-01
影响因子：
5.3
通讯作者：
ORKIN, SH
中科院分区：
文献类型：
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作者：
MERIKA, M;ORKIN, SH
GATA结合蛋白构成了一个转录因子家族，其识别符合共有WGATAR（W = A或T且R = A或G）的靶位点。在这里，我们已经使用的聚合酶链反应介导的随机位点选择的方法来评估在一个公正的方式的DNA结合特异性的加塔蛋白。与我们的预期相反，我们表明，加塔蛋白结合的各种图案，偏离先前分配的共识。许多非共有序列以高亲和力结合蛋白质，相当于常规加塔基序的亲和力。通过使用选定的序列作为探针在电泳迁移率变动分析，我们证明重叠，但不同的序列偏好加塔家族成员，指定其各自的DNA结合域。此外，我们提供了额外的证据的相互作用的氨基和羧基手指的加塔-1在定义其结合位点。通过进行共转染实验，我们还表明，反式激活平行的DNA结合。含有areA的指状结构域和加塔-1的激活结构域的嵌合蛋白能够通过加塔基序激活哺乳动物细胞中的转录。我们的研究结果表明，加塔蛋白可能选择性地调节基因表达的细胞中，他们是共表达的机制。
GATA-binding proteins constitute a family of transcription factors that recognize a target site conforming to the consensus WGATAR (W = A or T and R = A or G). Here we have used the method of polymerase chain reaction-mediated random site selection to assess in an unbiased manner the DNA-binding specificity of GATA proteins. Contrary to our expectations, we show that GATA proteins bind a variety of motifs that deviate from the previously assigned consensus. Many of the nonconsensus sequences bind protein with high affinity, equivalent to that of conventional GATA motifs. By using the selected sequences as probes in the electrophoretic mobility shift assay, we demonstrate overlapping, but distinct, sequence preferences for GATA family members, specified by their respective DNA-binding domains. Furthermore, we provide additional evidence for interaction of amino and carboxy fingers of GATA-1 in defining its binding site. By performing cotransfection experiments, we also show that transactivation parallels DNA binding. A chimeric protein containing the finger domain of areA and the activation domains of GATA-1 is capable of activating transcription in mammalian cells through GATA motifs. Our findings suggest a mechanism by which GATA proteins might selectively regulate gene expression in cells in which they are coexpressed.