Liu Ying, Zheng Wenlu, Zhang Wan, Chen Nan, Liu Yang, Chen Li, Zhou Xiaozhou, Chen Xingshuo, Zheng Haifeng, Li Xiaoyu
Key Laboratory of Bioorganic Chemistry and Molecular Engineering of the Ministry of Education , Beijing National Laboratory of Molecular Sciences , College of Chemistry and Molecular Engineering , Peking University , Beijing , China 100871 . Email:
Key Laboratory of Chemical Genomics , School of Chemical Biology and Biotechnology , Peking University Shenzhen Graduate School , Shenzhen , China 518055.
Chem Sci. 2015 Jan 1;6(1):745-751. doi: 10.1039/c4sc01953a. Epub 2014 Oct 1.
Characterization of transcription factor-DNA interaction is of high importance in elucidating the molecular mechanisms of gene transcriptions. DNA-based affinity probes were developed to capture and identify transcription factors by covalent crosslinking; however, the requirement of a crosslinker on the affinity probe remains a disadvantage, as the crosslinker itself often interferes with the protein-DNA interactions. We report a dual-probe method able to capture DNA-binding transcription factors with unmodified protein-binding sites in scenarios where conventional probes have failed. We have also shown the method's converse application in selecting specific transcription factor-binding DNA sequences from a probe library and its extension to studying proteins recognizing epigenetic marks. This study may provide a new tool for exploring DNA-binding proteins in biology.
转录因子与DNA相互作用的表征对于阐明基因转录的分子机制至关重要。基于DNA的亲和探针通过共价交联来捕获和鉴定转录因子；然而，亲和探针上需要交联剂仍然是一个缺点，因为交联剂本身常常会干扰蛋白质与DNA的相互作用。我们报道了一种双探针方法，在传统探针失效的情况下，该方法能够捕获具有未修饰蛋白质结合位点的DNA结合转录因子。我们还展示了该方法的反向应用，即从探针文库中选择特定的转录因子结合DNA序列，以及将其扩展到研究识别表观遗传标记的蛋白质。这项研究可能为探索生物学中的DNA结合蛋白提供一种新工具。
