An optogenetic tool for the activation of endogenous diaphanous-related formins induces thickening of stress fibers without an increase in contractility.
An optogenetic tool for the activation of endogenous diaphanous-related formins induces thickening of stress fibers without an increase in contractility.
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用于激活内源性透明相关福明的光遗传学工具可诱导应力纤维增厚而不增加收缩性。
DOI：
10.1002/cm.21115
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Zaidel-Bar,Ronen
中科院分区：
文献类型：
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作者：
Rao,MeghaVaman;Chu,Pei-Hsuan;Hahn,KlausMichael;Zaidel-Bar,Ronen
我们开发了一种光遗传技术来激活透明相关的福尔马林。我们的方法是基于燕麦光养蛋白1的光-氧-电压2结构域与从mDia1分离的透明自我调节结构域的融合。这种“关在笼子里”的透明自动调节域在黑暗中是不活跃的，但在蓝光存在时迅速激活内源性透明相关的福尔马林。使用F-肌动蛋白报告，我们观察到丝状足细胞和片状脂肪细胞的形成，以及沿现有应激纤维的F-肌动蛋白的稳定增加，从光激活后的几分钟内开始。有趣的是，我们没有观察到新的应力纤维的形成。值得注意的是，F-肌动蛋白1.9倍的增加并不能与应力纤维上肌球蛋白II的增加平行，而且根据局部粘连大小判断，纤维产生的张力量似乎没有变化。我们的结果表明在应激纤维中F-肌动蛋白的积累和收缩之间存在解偶联，并证明了可光激活的透明自调节结构域在细胞透明相关的Forin功能研究中的作用。©2013 Wiley期刊，Inc.
We have developed an optogenetic technique for the activation of diaphanous‐related formins. Our approach is based on fusion of the light‐oxygen‐voltage 2 domain ofAvena sativaPhototrophin1 to an isolated Diaphanous Autoregulatory Domain from mDia1. This “caged” diaphanous auto‐regulatory domain was inactive in the dark but in the presence of blue light rapidly activated endogenous diaphanous‐related formins. Using an F‐actin reporter, we observed filopodia and lamellipodia formation as well as a steady increase in F‐actin along existing stress fibers, starting within minutes of photo‐activation. Interestingly, we did not observe the formation of new stress fibers. Remarkably, a 1.9‐fold increase in F‐actin was not paralleled by an increase in myosin II along stress fibers and the amount of tension generated by the fibers, as judged by focal adhesion size, appeared unchanged. Our results suggest a decoupling between F‐actin accumulation and contractility in stress fibers and demonstrate the utility of photoactivatable diaphanous autoregulatory domain for the study of diaphanous‐related formin function in cells. © 2013 Wiley Periodicals, Inc.