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Ultraviolet-B-mediated induction of protein-protein interactions in mammalian cells

机译:紫外线B介导的哺乳动物细胞中蛋白质相互作用的诱导

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摘要

Light-sensitive proteins are useful tools to control protein localization, activation and gene expression, but are currently limited to excitation with red or blue light. Here we report a novel optogenetic system based on the ultraviolet-B-dependent interaction of the Arabidopsis ultraviolet-B photoreceptor UVR8 with COP1 that can be performed in visible light background. We use this system to induce nuclear accumulation of cytoplasmic green fluorescent protein fused to UVR8 in cells expressing nuclear COP1, and to recruit a nucleoplasmic red fluorescent protein fused to COP1 to chromatin in cells expressing UVR8-H2B. We also show that ultraviolet-B-dependent interactions between DNA-binding and transcription activation domains result in a linear induction of gene expression. The UVR8-COP1 interactions in mammalian cells can be induced using subsecond pulses of ultraviolet-B light and last several hours. As UVR8 photoperception is based on intrinsic tryptophan residues, these interactions do not depend on the addition of an exogenous chromophore.
机译:光敏蛋白是控制蛋白定位,激活和基因表达的有用工具,但目前仅限于红光或蓝光激发。在这里,我们报告了一种新型的光遗传系统,该系统基于拟南芥紫外线B感光体UVR8与COP1的紫外线B依赖性相互作用,可以在可见光背景下进行。我们使用该系统在表达核COP1的细胞中诱导与UVR8融合的胞质绿色荧光蛋白的核积累,并在表达UVR8-H2B的细胞中募集与COP1融合的核质红色荧光蛋白。我们还显示,DNA结合和转录激活域之间的依赖紫外线B的相互作用导致基因表达的线性诱导。可以使用亚秒级的紫外线-B脉冲诱导哺乳动物细胞中的UVR8-COP1相互作用,并持续数小时。由于UVR8的光感知是基于固有的色氨酸残基,因此这些相互作用不依赖于外源发色团的添加。

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