一种靶向 TUT4 的 LIN28:let-7 相互作用片段的纳米抗体可阻断 let-7 的尿嘧啶化。
A nanobody targeting the LIN28:let-7 interaction fragment of TUT4 blocks uridylation of let-7.
机构信息
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA 02115.
出版信息
Proc Natl Acad Sci U S A. 2020 Mar 3;117(9):4653-4663. doi: 10.1073/pnas.1919409117. Epub 2020 Feb 14.
The LIN28:pre-let-7:TUTase ternary complex regulates pluripotency and oncogenesis by controlling processing of the let-7 family of microRNAs. The complex oligouridylates the 3' ends of pre-let-7 molecules, leading to their degradation via the DIS3L2 exonuclease. Previous studies suggest that components of this complex are potential therapeutic targets in malignancies that aberrantly express LIN28. In this study we developed a functional epitope selection approach to identify nanobody inhibitors of the LIN28:pre-let-7:TUT4 complex. We demonstrate that one of the identified nanobodies, Nb-S2A4, targets the 106-residue LIN28:let-7 interaction (LLI) fragment of TUT4. Nb-S2A4 can effectively inhibit oligouridylation and monouridylation of pre-let-7g in vitro. Expressing Nb-S2A4 allows maturation of the let-7 species in cells expressing LIN28, highlighting the therapeutic potential of targeting the LLI fragment.
LIN28:pre-let-7:TUTase 三元复合物通过控制 let-7 微 RNA 家族的加工来调节多能性和致癌性。该复合物对 pre-let-7 分子的 3' 端进行寡聚化，导致它们通过 DIS3L2 核酸外切酶降解。先前的研究表明，该复合物的成分是异常表达 LIN28 的恶性肿瘤的潜在治疗靶点。在这项研究中，我们开发了一种功能表位选择方法，以鉴定针对 LIN28:pre-let-7:TUT4 复合物的纳米抗体抑制剂。我们证明，鉴定出的一种纳米抗体 Nb-S2A4 靶向 TUT4 的 106 个残基 LIN28:let-7 相互作用（LLI）片段。Nb-S2A4 可以有效抑制体外 pre-let-7g 的寡聚化和单聚化。表达 Nb-S2A4 允许在表达 LIN28 的细胞中 let-7 物种成熟，突出了靶向 LLI 片段的治疗潜力。
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