SELEX and missing phosphate contact analyses reveal flexibility within the AP-2α protein:DNA binding complex
SELEX and missing phosphate contact analyses reveal flexibility within the AP-2α protein:DNA binding complex
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DOI：
10.1093/nar/27.13.2760
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发表时间：
1999-07-01
影响因子：
14.9
通讯作者：
Williams, T
中科院分区：
文献类型：
--
作者：
Mohibullah, N;Donner, A;Williams, T
AP-2转录因子家族是由一个新的DNA结合域定义的，称为“基本螺旋-跨螺旋”基序。AP-2基因调节脊椎动物胚胎发生的重要方面，也与细胞增殖和肿瘤发生的控制有关，但AP-2蛋白控制的细胞靶点在很大程度上是不明确的。特别是，由于以前仅利用有限数量的序列来定义AP-2结合位点的性质，因此AP-2蛋白识别的DNA序列范围仍然未知。因此，我们利用SELEX分析来鉴定多个新的AP-2 α结合位点，此外，我们还设计了一种新的缺失磷酸盐和核苷酸竞争分析来表征AP-2 α蛋白所需的结合位点上的残基：DNA接触。这些研究表明，AP-2 α蛋白：DNA复合物是灵活的，并表明AP-2 α可以结合三个相关的序列基序：GCC N3 GGC， GCC N4 GGC和GCC N3/4 GGG，这些更精细的共识序列的可用性将有助于鉴定这一关键转录因子的靶基因。
The AP-2 family of transcription factors are defined by the presence of a novel DNA binding domain, termed a 'basic helix-span-helix' motif, The AP-2 genes regulate important aspects of vertebrate embryogenesis and have also been linked to the control of cell proliferation and tumorigenesis, but the cellular targets that the AP-2 proteins control are largely undefined. In particular, since only a limited number of sequences have previously been utilized to define the nature of the AP-2 binding site, the range of DNA sequences recognized by the AP-2 proteins remains unknown. We have therefore utilized a SELEX analysis to identify multiple new AP-2 alpha binding sites, Moreover, we have devised a novel missing phosphate and nucleotide competition analysis to characterize the residues in the binding site required for AP-2 alpha protein:DNA contact. These studies suggest that the AP-2 alpha protein:DNA complex is flexible and indicate that AP-2 alpha can bind three related sequence motifs: GCC N3 GGC, GCC N4 GGC and GCC N3/4 GGG, The availability of these more refined consensus sequences should assist in the identification of target genes for this critical transcription factor.