Department of Molecular Biology, University of Geneva, 30 quai Ernest-Ansermet, CH 1205 Geneva, Switzerland.
Nat Commun. 2013;4:1779. doi: 10.1038/ncomms2800.
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.
光敏感蛋白是控制蛋白质定位、激活和基因表达的有用工具,但目前仅限于用红光或蓝光进行激发。在这里,我们报告了一种基于拟南芥紫外光 B 型受体 UVR8 与 COP1 之间的紫外光 B 依赖性相互作用的新型光遗传学系统,该系统可以在可见光背景下进行。我们使用该系统诱导核表达 COP1 的细胞中细胞质绿色荧光蛋白融合到 UVR8 的核积累,并将核质红色荧光蛋白募集到表达 UVR8-H2B 的染色质上。我们还表明,DNA 结合和转录激活结构域之间的紫外光 B 依赖性相互作用导致基因表达的线性诱导。哺乳动物细胞中的 UVR8-COP1 相互作用可以使用亚秒级的紫外光脉冲诱导,持续数小时。由于 UVR8 的光感知是基于内在色氨酸残基,因此这些相互作用不依赖于外加发色团的添加。