Ultraviolet-B-mediated induction of protein-protein interactions in mammalian cells
Ultraviolet-B-mediated induction of protein-protein interactions in mammalian cells
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DOI：
10.1038/ncomms2800
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发表时间：
2013-04-01
影响因子：
16.6
通讯作者：
Halazonetis, Thanos D.
中科院分区：
文献类型：
--
作者：
Crefcoeur, Remco P.;Yin, Ruohe;Halazonetis, Thanos D.
光敏蛋白是控制蛋白定位、激活和基因表达的有用工具，但目前仅限于用红光或蓝光激发。在这里，我们报道了一种基于拟南芥紫外线 B 光感受器 UVR8 与 COP1 的紫外线 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。我们使用该系统在表达核 COP1 的细胞中诱导与 UVR8 融合的细胞质绿色荧光蛋白在核中积累，并在表达 UVR8-H2B 的细胞中将与 COP1 融合的核质红色荧光蛋白募集到染色质。我们还表明，DNA 结合域和转录激活域之间依赖于 UV-B 的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 UV-B 光的亚秒脉冲诱导，并持续几个小时。由于 UVR8 光感知基于内在色氨酸残基，因此这些相互作用不依赖于外源生色团的添加。
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.