Ballister Edward R, Aonbangkhen Chanat, Mayo Alyssa M, Lampson Michael A, Chenoweth David M
Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Department of Chemistry, School of Arts and Sciences, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Nat Commun. 2014 Nov 17;5:5475. doi: 10.1038/ncomms6475.
Regulated protein localization is critical for many cellular processes. Several techniques have been developed for experimental control over protein localization, including chemically induced and light-induced dimerization, which both provide temporal control. Light-induced dimerization offers the distinct advantage of spatial precision within subcellular length scales. A number of elegant systems have been reported that utilize natural light-sensitive proteins to induce dimerization via direct protein-protein binding interactions, but the application of these systems at cellular locations beyond the plasma membrane has been limited. Here we present a new technique to rapidly and reversibly control protein localization in living cells with subcellular spatial resolution using a cell-permeable, photoactivatable chemical inducer of dimerization. We demonstrate light-induced recruitment of a cytosolic protein to individual centromeres, kinetochores, mitochondria and centrosomes in human cells, indicating that our system is widely applicable to many cellular locations.
受调控的蛋白质定位对许多细胞过程至关重要。已经开发了几种用于实验控制蛋白质定位的技术,包括化学诱导和光诱导二聚化,这两种方法都能提供时间控制。光诱导二聚化在亚细胞长度尺度上具有空间精度的独特优势。已经报道了许多精巧的系统,它们利用天然的光敏感蛋白通过直接的蛋白质-蛋白质结合相互作用诱导二聚化,但这些系统在质膜以外的细胞位置的应用受到限制。在这里,我们提出了一种新技术,使用可透过细胞的、光可激活的二聚化化学诱导剂,以亚细胞空间分辨率在活细胞中快速且可逆地控制蛋白质定位。我们证明了在人类细胞中,光诱导胞质蛋白募集到单个着丝粒、动粒、线粒体和中心体,这表明我们的系统广泛适用于许多细胞位置。