NMR and EPR reveal a compaction of the RNA-binding protein FUS upon droplet formation
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Date
2021-05Type
- Journal Article
Citations
Cited 23 times in
Web of Science
Cited 23 times in
Scopus
ETH Bibliography
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Abstract
Many RNA-binding proteins undergo liquid–liquid phase separation, which underlies the formation of membraneless organelles, such as stress granules and P-bodies. Studies of the molecular mechanism of phase separation in vitro are hampered by the coalescence and sedimentation of organelle-sized droplets interacting with glass surfaces. Here, we demonstrate that liquid droplets of fused in sarcoma (FUS)—a protein found in cytoplasmic aggregates of amyotrophic lateral sclerosis and frontotemporal dementia patients—can be stabilized in vitro using an agarose hydrogel that acts as a cytoskeleton mimic. This allows their spectroscopic characterization by liquid-phase NMR and electron paramagnetic resonance spectroscopy. Protein signals from both dispersed and condensed phases can be observed simultaneously, and their respective proportions can be quantified precisely. Furthermore, the agarose hydrogel acts as a cryoprotectant during shock-freezing, which facilitates pulsed electron paramagnetic resonance measurements at cryogenic temperatures. Surprisingly, double electron–electron resonance measurements revealed a compaction of FUS in the condensed phase. Show more
Publication status
publishedExternal links
Journal / series
Nature Chemical BiologyVolume
Pages / Article No.
Publisher
Nature Publishing GroupOrganisational unit
03810 - Jeschke, Gunnar / Jeschke, Gunnar
08703 - Gruppe Klotzsch / Group Klotzsch
03591 - Allain, Frédéric / Allain, Frédéric
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Show all metadata
Citations
Cited 23 times in
Web of Science
Cited 23 times in
Scopus
ETH Bibliography
yes
Altmetrics