MCAK-mediated regulation of endothelial cell microtubule dynamics is mechanosensitive to myosin-II contractility
MCAK-mediated regulation of endothelial cell microtubule dynamics is mechanosensitive to myosin-II contractility
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DOI：
10.1091/mbc.e16-05-0306
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发表时间：
2017-05-01
影响因子：
3.3
通讯作者：
Myers, Kenneth A.
中科院分区：
文献类型：
--
作者：
D'Angelo, Lauren;Myer, Nicole M.;Myers, Kenneth A.
顺应性和维度机械感知是细胞感知细胞外基质(ECM)物理属性的过程，已知主要通过肌球蛋白II介导的肌动蛋白和微管(MT)细胞骨架的重组来驱动细胞分支和形状变化。亚细胞对MT动力学的调控是通过rac1-Aurora-A激酶通路在空间上控制的，该通路局部抑制有丝分裂着丝粒相关蛋白(MCAK)的MT解聚活性，从而促进MT的前沿生长和细胞极化。这些结果表明，MT生长动力学的调节与细胞与ECM的物理接触密切相关。在这里，我们验证了这样的假设：MCAK通过肌球蛋白II依赖的信号通路对MT生长动力学的顺应性和维度机械感觉介导的调控做出了贡献。我们将内皮细胞培养在胶原偶联的刚性或顺应性的聚丙烯酰胺内皮细胞上，以检测MCAK的表达对MT生长动力学和EC分支形态的影响。我们的结果表明，MCAK促进了在顺应性2D ECM上培养的ECs中MT的快速生长速度，但促进了在顺应性3D ECM上培养的ECs中MT的缓慢生长速度，并且这些作用是肌球蛋白II依赖的。此外，我们发现，3D ECM参与将MCAK介导的MT生长持续性调节与肌球蛋白II介导的生长持续性调节(特别是EC分支突起中的生长持续性调节)分开。
Compliance and dimensionality mechanosensing, the processes by which cells sense the physical attributes of the extracellular matrix (ECM), are known to drive cell branching and shape change largely through a myosin-II-mediated reorganization of the actin and microtubule (MT) cytoskeletons. Subcellular regulation of MT dynamics is spatially controlled through a Rac1-Aurora-A kinase pathway that locally inhibits the MT depolymerizing activity of mitotic centromere-associated kinesin (MCAK), thereby promoting leading-edge MT growth and cell polarization. These results suggest that the regulation of MT growth dynamics is intimately linked to physical engagement of the cell with the ECM. Here, we tested the hypothesis that MCAK contributes to compliance and dimensionality mechanosensing-mediated regulation of MT growth dynamics through a myosin-II-dependent signaling pathway. We cultured endothelial cells (ECs) on collagen-coupled stiff or compliant polyacrylamide ECMs to examine the effects of MCAK expression on MT growth dynamics and EC branching morphology. Our results identify that MCAK promotes fast MT growth speeds in ECs cultured on compliant 2D ECMs but promotes slow MT growth speeds in ECs cultured on compliant 3D ECMs, and these effects are myosin-II dependent. Furthermore, we find that 3D ECM engagement uncouples MCAK-mediated regulation of MT growth persistence from myosin-II-mediated regulation of growth persistence specifically within EC branched protrusions.