Direct Observation and Analysis of the Dynamics of the Photoresponsive Transcription Factor GAL4.
Direct Observation and Analysis of the Dynamics of the Photoresponsive Transcription Factor GAL4.
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DOI：
10.1002/anie.201900610
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发表时间：
2019-04
影响因子：
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通讯作者：
G. Raghavan;Kumi Hidaka;H. Sugiyama;Masayuki Endo
中科院分区：
文献类型：
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作者：
G. Raghavan;Kumi Hidaka;H. Sugiyama;Masayuki Endo
本文报道了利用高速原子力显微镜（HS-AFM）直接观察光响应性转录因子融合体GAL 4-VVD和DNA之间的动态相互作用。观察到一系列不同的GAL 4-VVD运动，如结合、滑动、停滞和解离。还观察到两个双链（ds）DNA上的链间跳跃。使用含有5个GAL 4结合位点的长底物DNA链进行的详细分析表明，GAL 4-VVD在dsDNA上通过滑动和跳跃随机移动以快速找到特异性结合位点，然后停滞在特异性位点以形成稳定的复合物形成。这些结果表明，存在不同的构象的蛋白质，使滑动和失速。这种具有纳米级分辨率的单分子成像系统提供了对DNA结合蛋白所使用的搜索机制的深入了解。
Herein, the direct visualization of the dynamic interaction between a photoresponsive transcription factor fusion, GAL4-VVD, and DNA using high-speed atomic force microscopy (HS-AFM) is reported. A series of different GAL4-VVD movements, such as binding, sliding, stalling, and dissociation, was observed. Inter-strand jumping on two double-stranded (ds) DNAs was also observed. Detailed analysis using a long substrate DNA strand containing five GAL4-binding sites revealed that GAL4-VVD randomly moved on the dsDNA using sliding and hopping to rapidly find specific binding sites, and then stalled to the specific sites to form a stable complex formation. These results suggest the existence of different conformations of the protein to enable sliding and stalling. This single-molecule imaging system with nanoscale resolution provides an insight into the searching mechanism used by DNA-binding proteins.