Spatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets.

TitleSpatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets.
Publication TypeJournal Article
Year of Publication2017
AuthorsShin, Y, Berry, J, Pannucci, N, Haataja, MP, Toettcher, JE, Brangwynne, CP
JournalCell
Volume168
Issue1-2
Pagination159-171.e14
Date Published2017 Jan 12
ISSN1097-4172
KeywordsAnimals, Arabidopsis Proteins, Cryptochromes, Intrinsically Disordered Proteins, Kinetics, Light, Mice, Models, Chemical, Molecular Imaging, NIH 3T3 Cells, Optogenetics, Phase Transition, Protein Interaction Maps, Proteins
Abstract

<p>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 that 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 that undergo aging into irreversible aggregates. This system can thus elucidate not only physiological phase transitions but also their link to pathological aggregates.</p>

DOI10.1016/j.cell.2016.11.054
Alternate JournalCell
PubMed ID28041848
PubMed Central IDPMC5562165
Grant ListDP2 EB024247 / EB / NIBIB NIH HHS / United States
DP2 GM105437 / GM / NIGMS NIH HHS / United States
U01 DA040601 / DA / NIDA NIH HHS / United States