小分子抑制剂破坏 let-7 寡核苷酸化并释放 LIN28 对 let-7 加工的选择性阻断。
Small-Molecule Inhibitors Disrupt let-7 Oligouridylation and Release the Selective Blockade of let-7 Processing by LIN28.
机构信息
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA.
Stem Cell Program, Boston Children's Hospital, Boston, MA, USA; Department of Pediatric Hematology/Oncology, Boston Children's Hospital and Dana-Farber Cancer Institute, Boston, MA, USA.
出版信息
Cell Rep. 2018 Jun 5;23(10):3091-3101. doi: 10.1016/j.celrep.2018.04.116.
LIN28 is an RNA-binding protein that regulates the maturation of the let-7 family of microRNAs by bipartite interactions with let-7 precursors through its two distinct cold shock and zinc-knuckle domains. Through inhibition of let-7 biogenesis, LIN28 functions as a pluripotency factor, as well as a driver of tumorigenesis. Here, we report a fluorescence polarization assay to identify small-molecule inhibitors for both domains of LIN28 involved in let-7 interactions. Of 101,017 compounds screened, six inhibit LIN28:let-7 binding and impair LIN28-mediated let-7 oligouridylation. Upon further characterization, we demonstrate that the LIN28 inhibitor TPEN destabilizes the zinc-knuckle domain of LIN28, while LI71 binds the cold shock domain to suppress LIN28's activity against let-7 in leukemia cells and embryonic stem cells. Our results demonstrate selective pharmacologic inhibition of individual domains of LIN28 and provide a foundation for therapeutic inhibition of the let-7 biogenesis pathway in LIN28-driven diseases.
LIN28 是一种 RNA 结合蛋白，通过其两个独特的冷休克和锌指结构域与 let-7 前体的二分相互作用，调节 let-7 家族 microRNA 的成熟。通过抑制 let-7 的生物发生，LIN28 作为多能性因子以及肿瘤发生的驱动因子发挥作用。在这里，我们报告了一种荧光偏振测定法，以鉴定参与 let-7 相互作用的 LIN28 的两个结构域的小分子抑制剂。在筛选了 101017 种化合物后，有 6 种抑制 LIN28：let-7 结合并损害 LIN28 介导的 let-7 寡聚化。经过进一步的表征，我们证明 LIN28 抑制剂 TPEN 使 LIN28 的锌指结构域不稳定，而 LI71 结合冷休克结构域以抑制 LIN28 在白血病细胞和胚胎干细胞中对 let-7 的活性。我们的结果证明了对 LIN28 单个结构域的选择性药理抑制，并为 LIN28 驱动疾病中 let-7 生物发生途径的治疗性抑制提供了基础。
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