Heterogeneous dynamics in DNA site discrimination by the structurally homologous DNA-binding domains of ETS-family transcription factors.
Heterogeneous dynamics in DNA site discrimination by the structurally homologous DNA-binding domains of ETS-family transcription factors.
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DOI：
10.1093/nar/gkv267
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发表时间：
2015-04-30
影响因子：
14.9
通讯作者：
Poon GM
中科院分区：
文献类型：
--
作者：
He G;Tolic A;Bashkin JK;Poon GM
ETS家族的转录因子证实了目前在具有重叠DNA序列偏好的真核遗传调节因子如何实现靶位点特异性方面的不确定性。PU.1和Ets-1代表了研究ETS蛋白的位点识别的原型，因为它们的DNA结合结构域在序列上是最不同的，但它们具有显著的重叠DNA结合结构。为了深入了解这两种蛋白质的DNA识别的热力学和动力学对比，我们研究了它们的DNA结合域的结构和动力学。电泳迁移率的复合物形成的两个同系物与循环排列的结合位点表现出显着的动态差异，只有DNA复合物的PU。1。通过动态光散射的自由溶液测量表明PU.1比Ets-1更具动态性;此外，动态变化强烈耦合到PU.1，但不是Ets-1的网站歧视。通过DNA足迹法对蛋白质/DNA界面进行的询问显示，PU.1/DNA和Ets-1/DNA复合物对硫酸二甲酯的可及性相似，表明PU.1/DNA复合物的动力学主要位于该界面之外。一个基于信息的分析两个同源物的结合基序表明，在PU.1的能力，执行一个更严格的序列偏好比Ets-1和它的近端序列同源物的动态耦合的作用。
The ETS family of transcription factors exemplifies current uncertainty in how eukaryotic genetic regulators with overlapping DNA sequence preferences achieve target site specificity. PU.1 and Ets-1 represent archetypes for studying site discrimination by ETS proteins because their DNA-binding domains are the most divergent in sequence, yet they share remarkably superimposable DNA-bound structures. To gain insight into the contrasting thermodynamics and kinetics of DNA recognition by these two proteins, we investigated the structure and dynamics of site discrimination by their DNA-binding domains. Electrophoretic mobilities of complexes formed by the two homologs with circularly permuted binding sites showed significant dynamic differences only for DNA complexes of PU.1. Free solution measurements by dynamic light scattering showed PU.1 to be more dynamic than Ets-1; moreover, dynamic changes are strongly coupled to site discrimination by PU.1, but not Ets-1. Interrogation of the protein/DNA interface by DNA footprinting showed similar accessibility to dimethyl sulfate for PU.1/DNA and Ets-1/DNA complexes, indicating that the dynamics of PU.1/DNA complexes reside primarily outside that interface. An information-based analysis of the two homologs’ binding motifs suggests a role for dynamic coupling in PU.1's ability to enforce a more stringent sequence preference than Ets-1 and its proximal sequence homologs.
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影响因子：
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作者：
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通讯作者：
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