Patterning Microtubule Network Organization Reshapes Cell-Like Compartments.
Patterning Microtubule Network Organization Reshapes Cell-Like Compartments.
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DOI：
10.1021/acssynbio.0c00575
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发表时间：
2021-06-18
影响因子：
4.7
通讯作者：
Good, Matthew C.
中科院分区：
文献类型：
--
作者：
Bermudez, Jessica G.;Deiters, Alexander;Good, Matthew C.
关键词：
真核细胞含有由动态微管丝组成的细胞骨架网络，其空间组织是高度可塑的。专门的微管结构针对不同的细胞类型进行优化，并随着振荡细胞周期进行重塑。这些空间上不同的微管网络被认为是由微管调节剂和马达的活性和定位引起的，并且进一步由来自细胞边界的物理力形成。鉴于活细胞的复杂性和冗余性，将微管网络组织的生物化学和物理贡献分开是具有挑战性的。因此，我们试图开发一种最小的细胞样系统，以实时操纵和空间模式化细胞骨架组分的组织，提供一个机会来建立不同的空间结构，并确定它们如何被细胞边界塑造或重塑。我们构建了一个系统，用于在细胞大小的乳液隔室中诱导蛋白质组分的空间图案化，并使用它来实时驱动微管网络组织。我们使用小分子和光控制动态蛋白质重新定位，并减缓脂质单层内的横向扩散，以利用聚焦照明创建稳定的微图案。通过将微管相互作用蛋白融合到光化学二聚化结构域，我们指导了微管网络的空间组织。聚合微管的皮层模式导致对称性破坏和力量，显着重塑隔间。我们的系统在细胞生物学中的应用，以表征生物化学成分和物理边界条件对微管网络组织的贡献。此外，主动形状控制在原细胞工程和增强合成细胞的功能中有用途。
Eukaryotic cells contain a cytoskeletal network comprised of dynamic microtubule filaments whose spatial organization is highly plastic. Specialized microtubule architectures are optimized for different cell types and remodel with the oscillatory cell cycle. These spatially distinct microtubule networks are thought to arise from the activity and localization of microtubule regulators and motors and are further shaped by physical forces from the cell boundary. Given complexities and redundancies of a living cell, it is challenging to disentangle the separate biochemical and physical contributions to microtubule network organization. Therefore, we sought to develop a minimal cell-like system to manipulate and spatially pattern the organization of cytoskeletal components in real-time, providing an opportunity to build distinct spatial structures and to determine how they are shaped by or reshape cell boundaries. We constructed a system for induced spatial patterning of protein components within cell-sized emulsion compartments and used it to drive microtubule network organization in real-time. We controlled dynamic protein relocalization using small molecules and light and slowed lateral diffusion within the lipid monolayer to create stable micropatterns with focused illumination. By fusing microtubule interacting proteins to optochemical dimerization domains we directed the spatial organization of microtubule networks. Cortical patterning of polymerizing microtubules leads to symmetry breaking and forces that dramatically reshapes the compartment. Our system has applications in cell biology to characterize the contributions of biochemical components and physical boundary conditions to microtubule network organization. Additionally, active shape control has uses in protocell engineering and for augmenting the functionalities of synthetic cells.
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DOI：
10.1073/pnas.1222097110
发表时间：
2013-07-23
影响因子：
11.1
作者：
Di Rienzo, Carmine;Gratton, Enrico;Cardarelli, Francesco
通讯作者：
Cardarelli, Francesco
影响因子：
4.6
作者：
Akhunzada, Muhammad Jan;D'Autilia, Francesca;Brancato, Giuseppe
通讯作者：
Brancato, Giuseppe
影响因子：
11.6
作者：
Dhonukshe, P;Gadella, TWJ
通讯作者：
Gadella, TWJ
影响因子：
3.5
作者：
Cappelletti, G;Surrey, T;Maci, R
通讯作者：
Maci, R
影响因子：
4.7
作者：
Hallett RA;Zimmerman SP;Yumerefendi H;Bear JE;Kuhlman B
通讯作者：
Kuhlman B