A Phytochrome-Derived Photoswitch for Intracellular Transport
A Phytochrome-Derived Photoswitch for Intracellular Transport
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DOI：
10.1021/acssynbio.6b00333
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发表时间：
2017-07-01
影响因子：
4.7
通讯作者：
Kapitein, Lukas C.
中科院分区：
文献类型：
--
作者：
Adrian, Max;Nijenhuis, Wilco;Kapitein, Lukas C.
细胞依赖于细胞器的适当位置，这表明细胞器位置的主动操纵可以用于探索空间细胞生物学和修复细胞器错位造成的细胞缺陷。最近，蓝光依赖于特定马达到选定细胞器的募集被证明可以改变细胞器的运动和定位，但这些方法缺乏快速和主动的可逆性。光敏色素B与其相互作用因子的光依赖性相互作用表现为光开关作用，在红光下发生二聚化，在远红光下发生解离。在这里，我们将光敏色素结构域改造成光开关，用于细胞内运输，使细胞器和马达蛋白之间能够发生可逆的相互作用。使用图案化照明和活细胞成像，我们证明了该系统提供了前所未有的时空控制。我们还证明，它可以与蓝光依赖系统结合使用，以独立控制两个不同细胞器的定位。对细胞器运动和定位的精确光遗传控制将为更好地理解和控制细胞的空间生物学提供依据。
Cells depend on the proper positioning of their organelles, suggesting that active manipulation of organelle positions can be used to explore spatial cell biology and to restore cellular defects caused by organelle misplacement. Recently, blue light dependent recruitment of specific motors to selected organelles has been shown to alter organelle motility and positioning, but these approaches lack rapid and active reversibility. The light-dependent interaction of phytochrome B with its interacting factors has been shown to function as a photoswitch, dimerizing under red light and dissociating under far-red light. Here we engineer phytochrome domains into photoswitches for intracellular transport that enable the reversible interaction between organelles and motor proteins. Using patterned illumination and live-cell imaging, we demonstrate that this system provides unprecedented spatiotemporal control. We also demonstrate that it can be used in combination with a blue-light dependent system to independently control the positioning of two different organelles. Precise optogenetic control of organelle motility and positioning will provide a better understianding of and control over the spatial biology of cells.