The disordered DNA-binding domain of p53 is indispensable for forming an encounter complex to and jumping along DNA
The disordered DNA-binding domain of p53 is indispensable for forming an encounter complex to and jumping along DNA
复制标题
p53 无序的 DNA 结合域对于形成与 DNA 的相遇复合物并沿着 DNA 跳跃是必不可少的
DOI：
10.1016/j.bbrc.2020.12.006
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发表时间：
2021
影响因子：
3.1
通讯作者：
Kamagata Kiyoto
中科院分区：
文献类型：
--
作者：
Graha Subekti Dwiky Rendra;Kamagata Kiyoto
肿瘤抑制因子p53利用易化扩散机制来搜索并结合靶DNA序列。亚毫秒单分子荧光跟踪表明，p53形成一个短暂的遭遇复合物的DNA，然后转换为长寿命的复合物，可以移动和跳跃沿着DNA在目标搜索。为了揭示p53的每个DNA结合结构域在这些过程中的作用，我们研究了两个p53突变体缺乏两个DNA结合结构域中的任何一个;结构化的核心和无序的C-末端结构域，使用亚毫秒单分子荧光显微镜。我们发现，C-末端结构域是所需的遭遇复杂的形成和转换为长寿命的复杂。长寿命复合物由核心结构域以及C-末端结构域稳定。此外，在高盐浓度下，只有C-末端结构域参与p53沿着DNA的跳跃。我们认为p53的柔性C端结构域是围绕DNA的，它可以形成相遇复合物，转化为长寿命复合物，并使p53在跳跃后能够降落在DNA上。
The tumor suppressor p53 utilizes a facilitated diffusion mechanism to search for and bind to target DNA sequences. Sub-millisecond single-molecule fluorescence tracking demonstrated that p53 forms a short-lived encounter complex to DNA then converts to the long-lived complex that can move and jump along DNA during the target search. To reveal the role of each DNA-binding domain of p53 in these processes, we investigated two p53 mutants lacking either of two DNA-binding domains; structured core and disordered C-terminal domains, using sub-millisecond single-molecule fluorescence microscopy. We found that the C-terminal domain is required for the encounter complex formation and conversion to the long-lived complex. The long-lived complex is stabilized by the core domain as well as the C-terminal domain. Furthermore, only the C-terminal domain participates in the jump of p53 along DNA at a high salt concentration. We propose that the flexible C-terminal domain of p53 is twined around DNA, which can form the encounter complex, convert to the long-lived complex, and enable p53 to land on DNA after the jump.