Regulatable assembly of synthetic microtubule architectures using engineered MAP-IDR condensates.
Regulatable assembly of synthetic microtubule architectures using engineered MAP-IDR condensates.
复制标题
使用工程 MAP-IDR 冷凝物可调节合成微管结构的组装。
DOI：
10.1101/2023.03.14.532644
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Coyle,ScottM
中科院分区：
文献类型：
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作者：
Chang,Chih-Chia;Coyle,ScottM
微管细丝利用微管相关蛋白(MAP)组装成对细胞过程至关重要的高阶结构和机器。然而，指导细胞中新结构形成的合成图谱的设计是具有挑战性的，因为纳米级的生化活动必须跨越微米级的长度来组织。在这里，我们开发了合成MAP-IDR缩合物(SynMAP)，它在体外和哺乳动物细胞中提供可调节和可调节的高阶微管结构组装。SynMAP利用来自少突胶质细胞的一个小的微管结合域(TPPP)，其活性可以通过与凝聚形成的IDR序列相互作用而被合成地重新连接。这种结合使SynMAP能够自组织多价结构，将微管连接并连接到合成结构中。调节微管结合和凝聚形成组件之间的连接允许SynMAP作为更复杂的细胞骨架电路中的节点，其中微管结构的形成和动力学可以由小分子或细胞信号输入控制。通过系统地测试一组SynMAP电路设计，我们定义了一种纳米级(通过微管结合)和微级(通过凝聚体形成)的微管结构动态组装的两级控制方案。SynMAP为设计更复杂的微管结构和细胞机器提供了一个紧凑和合理的可工程化起点。
Microtubules filaments are assembled into higher-order structures and machines critical for cellular processes using microtubule-associated proteins (MAPs). However, the design of synthetic MAPs that direct the formation of new structures in cells is challenging, as nanoscale biochemical activities must be organized across micron length-scales. Here we develop synthetic MAP-IDR condensates (synMAPs) that provide tunable and regulatable assembly of higher-order microtubule structures in vitro and in mammalian cells. synMAPs harness a small microtubule-binding domain from oligodendrocytes (TPPP) whose activity can be synthetically rewired by interaction with condensate-forming IDR sequences. This combination allows synMAPs to self-organize multivalent structures that bind and bridge microtubules into synthetic architectures. Regulating the connection between the microtubule-binding and condensate-forming components allows synMAPs to act as nodes in more complex cytoskeletal circuits in which the formation and dynamics of the microtubule structure can be controlled by small molecules or cell-signaling inputs. By systematically testing a panel of synMAP circuit designs, we define a two-level control scheme for dynamic assembly of microtubule architectures at the nanoscale (via microtubule-binding) and microscale (via condensate formation). synMAPs provide a compact and rationally engineerable starting point for the design of more complex microtubule architectures and cellular machines.