Department of Pharmacology, University of Colorado School of Medicine, Aurora, CO, USA.
Nat Commun. 2021 Mar 22;12(1):1809. doi: 10.1038/s41467-021-22096-1.
Dynamic membraneless compartments formed by protein condensates have multifunctional roles in cellular biology. Tools that inducibly trigger condensate formation have been useful for exploring their cellular function, however, there are few tools that provide inducible control over condensate disruption. To address this need we developed DisCo (Disassembly of Condensates), which relies on the use of chemical dimerizers to inducibly recruit a ligand to the condensate-forming protein, triggering condensate dissociation. We demonstrate use of DisCo to disrupt condensates of FUS, associated with amyotrophic lateral sclerosis, and to prevent formation of polyglutamine-containing huntingtin condensates, associated with Huntington's disease. In addition, we combined DisCo with a tool to induce condensates with light, CRY2olig, achieving bidirectional control of condensate formation and disassembly using orthogonal inputs of light and rapamycin. Our results demonstrate a method to manipulate condensate states that will have broad utility, enabling better understanding of the biological role of condensates in health and disease.
由蛋白质凝聚物形成的动态无膜隔室在细胞生物学中具有多种功能。能够诱导凝聚物形成的工具对于探索其细胞功能非常有用，但是，很少有工具可以对凝聚物的破坏进行诱导控制。为了解决这一需求，我们开发了 DisCo（凝聚物的解体），它依赖于使用化学二聚体来诱导募集配体到形成凝聚物的蛋白质上，从而触发凝聚物解离。我们证明了 DisCo 可用于破坏与肌萎缩性侧索硬化症相关的 FUS 凝聚物，并防止与亨廷顿病相关的多聚谷氨酰胺含 huntingtin 凝聚物的形成。此外，我们将 DisCo 与一种用光诱导凝聚物的工具 CRY2olig 结合使用，使用光和雷帕霉素的正交输入实现了凝聚物形成和拆卸的双向控制。我们的结果证明了一种操纵凝聚物状态的方法，该方法具有广泛的用途，能够更好地理解凝聚物在健康和疾病中的生物学作用。
