Institute of Organic Chemistry and Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, Innrain 80/82, 6020, Innsbruck, Austria.
Boston Children's Hospital, Harvard Medical School, Boston, MA, 02115, USA.
Nat Commun. 2020 Nov 13;11(1):5750. doi: 10.1038/s41467-020-19144-7.
Nuclear export complexes composed of rev response element (RRE) ribonucleic acid (RNA) and multiple molecules of rev protein are promising targets for the development of therapeutic strategies against human immunodeficiency virus type 1 (HIV-1), but their assembly remains poorly understood. Using native mass spectrometry, we show here that rev initially binds to the upper stem of RRE IIB, from where it is relayed to binding sites that allow for rev dimerization. The newly discovered binding region implies initial rev recognition by nucleotides that are not part of the internal loop of RRE stem IIB RNA, which was previously identified as the preferred binding region. Our study highlights the unique capability of native mass spectrometry to separately study the binding interfaces of RNA/protein complexes of different stoichiometry, and provides a detailed understanding of the mechanism of RRE/rev association with implications for the rational design of potential drugs against HIV-1 infection.
核输出复合物由 rev 反应元件 (RRE) RNA 和多个 rev 蛋白分子组成，是开发针对人类免疫缺陷病毒 1 型 (HIV-1) 的治疗策略的有希望的靶点，但它们的组装仍知之甚少。本文使用天然质谱法表明，rev 最初与 RRE IIB 的上部茎结合，然后从那里传递到允许 rev 二聚化的结合位点。新发现的结合区域意味着 rev 最初识别的核苷酸不是 RRE 茎 IIB RNA 内部环的一部分，以前认为该区域是首选结合区域。我们的研究强调了天然质谱法分别研究不同化学计量的 RNA/蛋白质复合物结合界面的独特能力，并提供了对 RRE/rev 与 HIV-1 感染潜在药物合理设计相关的关联机制的详细了解。
