PASTEUR, Département de chimie, École normale supérieure, Paris Sciences et Lettres (PSL) University, Sorbonne Université, CNRS, 75005 Paris, France.
Institute for Integrated Cell-Material Sciences, Kyoto University, 606-8501 Kyoto, Japan.
Proc Natl Acad Sci U S A. 2019 Jul 2;116(27):13346-13351. doi: 10.1073/pnas.1900310116. Epub 2019 Jun 19.
The spatial regulation of messenger RNA (mRNA) translation is central to cellular functions and relies on numerous complex processes. Biomimetic approaches could bypass these endogenous complex processes, improve our comprehension of the regulation, and allow for controlling local translation regulations and functions. However, the causality between local translation and nascent protein function remains elusive. Here, we developed a nanoparticle (NP)-based strategy to magnetically control mRNA spatial patterns in mammalian cell extracts and investigate how local translation impacts nascent protein localization and function. By monitoring the translation of the magnetically localized mRNAs, we show that mRNA-NP complexes operate as a source for the continuous production of proteins from defined positions. By applying this approach to actin-binding proteins, we triggered the local formation of actin cytoskeletons and identified the minimal requirements for spatial control of the actin filament network. In addition, our bottom-up approach identified a role for mRNA as a translation-coupled scaffold for the function of nascent N-terminal protein domains. Our approach will serve as a platform for regulating mRNA localization and investigating the function of nascent protein domains during translation.
信使 RNA（mRNA）翻译的空间调控是细胞功能的核心，依赖于许多复杂的过程。仿生方法可以绕过这些内源性的复杂过程，增进我们对调控的理解，并允许控制局部翻译调控和功能。然而，局部翻译和新生蛋白质功能之间的因果关系仍然难以捉摸。在这里，我们开发了一种基于纳米颗粒（NP）的策略，在哺乳动物细胞提取物中控制 mRNA 的空间模式，并研究局部翻译如何影响新生蛋白质的定位和功能。通过监测磁定位的 mRNA 的翻译，我们表明 mRNA-NP 复合物作为从特定位置连续产生蛋白质的来源。通过将这种方法应用于肌动蛋白结合蛋白，我们触发了肌动蛋白细胞骨架的局部形成，并确定了空间控制肌动蛋白丝网络的最小要求。此外，我们的自下而上的方法确定了 mRNA 作为新生 N 端蛋白结构域功能的翻译偶联支架的作用。我们的方法将作为调节 mRNA 定位和研究翻译过程中新生蛋白结构域功能的平台。
