Spatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets.
Spatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets.
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DOI：
10.1016/j.cell.2016.11.054
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发表时间：
2017-01-12
期刊：
影响因子：
64.5
通讯作者：
Brangwynne CP
中科院分区：
文献类型：
--
作者：
Shin Y;Berry J;Pannucci N;Haataja MP;Toettcher JE;Brangwynne CP
由内在无序蛋白质区域（IDR）驱动的相变已经成为组装液体样RNA/蛋白质（RNP）体和其他无膜细胞器的普遍机制。然而，缺乏控制细胞内相变的工具限制了我们理解它们在细胞生理学和疾病中的作用的能力。在这里，我们介绍了一个光遗传学平台，它使用光来激活活细胞中IDR介导的相变。我们使用这个“optoDroplet”系统来研究由各种RNP体蛋白（包括FUS、DDX 4和HNRNPA 1）的IDR驱动的凝聚相。在浓度阈值以上，这些构建体经历光激活相分离，形成时空可定义的液体optoDroplets。FUS optoDroplet组装即使在多次激活循环后也是完全可逆的。然而，被驱入相界深处的细胞形成固体状凝胶，其经历老化成不可逆的聚集体。因此，该系统不仅可以阐明生理相变，而且还可以阐明它们与病理聚集体的联系。光依赖性触发蛋白质缔合允许选择性地时间和空间控制液滴和凝胶形成，旨在了解细胞内不同形式的无膜体和纤维状结构。
Phase transitions driven by intrinsically disordered protein regions (IDRs) have emerged as a ubiquitous mechanism for assembling liquid-like RNA/protein (RNP) bodies and other membrane-less organelles. However, a lack of tools to control intracellular phase transitions limits our ability to understand their role in cell physiology and disease. Here, we introduce an optogenetic platform, which uses light to activate IDR-mediated phase transitions in living cells. We use this “optoDroplet” system to study condensed phases driven by the IDRs of various RNP body proteins, including FUS, DDX4, and HNRNPA1. Above a concentration threshold, these constructs undergo light-activated phase separation, forming spatiotemporally-definable liquid optoDroplets. FUS optoDroplet assembly is fully reversible even after multiple activation cycles. However, cells driven deep within the phase boundary form solid-like gels, which undergo aging into irreversible aggregates. This system can thus elucidate not only physiological phase transitions, but also their link to pathological aggregates. Light-dependent triggering of protein association allows selective temporal and spatial control of droplet and gel formation aimed at understanding the different forms of membrane-less bodies and fibrillar structures within cells.
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影响因子：
48
作者：
Kennedy, Matthew J.;Hughes, Robert M.;Peteya, Leslie A.;Schwartz, Joel W.;Ehlers, Michael D.;Tucker, Chandra L.
通讯作者：
Tucker, Chandra L.
影响因子：
8.6
作者：
Lee, Chiu Fan;Brangwynne, Clifford P.;Juelicher, Frank
通讯作者：
Juelicher, Frank
影响因子：
2.9
作者：
LANGER, JS;SCHWARTZ, AJ
通讯作者：
SCHWARTZ, AJ
影响因子：
8.6
作者：
Cardinaux, Frederic;Gibaud, Thomas;Schurtenberger, Peter
通讯作者：
Schurtenberger, Peter
DOI：
10.1073/pnas.1509317112
发表时间：
2015-09-22
影响因子：
11.1
作者：
Berry, Joel;Weber, Stephanie C.;Brangwynne, Clifford P.
通讯作者：
Brangwynne, Clifford P.