PDF(Pubmed)
Abstract：
Within a shared cytoplasm, filamentous actin (F-actin) plays numerous and critical roles across the cell body. Cells rely on actin-binding proteins (ABPs) to organize F-actin and to integrate its polymeric characteristics into diverse cellular processes. Yet, the multitude of ABPs that engage with and shape F-actin make studying a single ABP\'s influence on cellular activities a significant challenge. Moreover, without a means of manipulating actin-binding subcellularly, harnessing the F-actin cytoskeleton for synthetic biology purposes remains elusive. Here, we describe a suite of designed proteins, Controllable Actin-binding Switch Tools (CASTs), whose actin-binding behavior can be controlled with external stimuli. CASTs were developed that respond to different external inputs, providing options for turn-on kinetics and enabling orthogonality and multiplexing. Being genetically encoded, we show that CASTs can be inserted into native protein sequences to control F-actin association locally and engineered into structures to control cell and tissue shape and behavior.
摘要：
在共享的细胞质中，丝状肌动蛋白（F-actin）在整个细胞体中起着许多关键的作用。细胞依赖于肌动蛋白结合蛋白（ABP）来组织F-肌动蛋白并将其聚合特征整合到不同的细胞过程中。然而，与F-肌动蛋白接触和形成的大量ABP使研究单个ABP对细胞活动的影响成为一项重大挑战。此外，没有操纵肌动蛋白结合亚细胞的手段，利用F-肌动蛋白细胞骨架用于合成生物学目的仍然难以捉摸。这里,我们描述了一套设计的蛋白质，可控肌动蛋白结合开关工具（CAST），其肌动蛋白结合行为可以用外部刺激控制。CAST被开发出来，以响应不同的外部输入，提供开启动力学选项并实现正交性和多路复用。被基因编码，我们表明，CAST可以插入到天然蛋白质序列中，以控制F-肌动蛋白结合局部和工程结构，以控制细胞和组织的形状和行为。
