Residue-by-residue view of in vitro FUS granules that bind the C-terminal domain of RNA polymerase II

By K. Burke, Abigail M. Janke, Christy L Rhine, Nicolas L. Fawzi. Molecules and Cells, 2015

814 citations recorded by the source corpus.

SUMMARY Phase-separated states of proteins underlie ribonucleoprotein (RNP) granules and nuclear RNA-binding protein assemblies that may nucleate protein inclusions associated with neurodegenerative diseases. We report that the N-terminal low complexity domain of the RNA-binding protein Fused in Sarcoma (FUS LC) is structurally disordered and forms a liquid-like phase-separated state resembling RNP granules. This state directly binds the C-terminal domain of RNA polymerase II. Phase-separated FUS lacks static structures as probed by fluorescence microscopy, indicating they are distinct from both protein inclusions and hydrogels. We use solution nuclear magnetic resonance spectroscopy to directly probe the dynamic architecture within FUS liquid phase-separated assemblies. Importantly, we find that FUS LC retains disordered secondary structure even in the liquid phase-separated state. Therefore, we propose that disordered protein granules, even those made of aggregation-prone prion-like domains, are dynamic and disordered molecular assemblies with transiently formed protein-protein contacts.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/a4596280f506f89e8380734fbd0c6ddbabf0862c

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Citations in the corpus

Sequence Determinants of Intracellular Phase Separation by Complex Coacervation of a Disordered Protein (2016, background): “For several of these, Tyr residues promote phase separation or recruitment into phase-separated structures (Burke et al., 2015; Jiang et al., 2015; Kato et al., 2012; Molliex et al., 2015; Patel et al., 2015).”
Spatiotemporal control of intracellular phase transitions using light-activated optoDroplets (2016, positive): “While the self-associating N-terminal IDR of FUS has been shown to be necessary and sufficient for liquid-liquid phase separation (Patel et al., 2015), C-terminal RNA binding domains appear to further promote phase separation ((Burke et al., 2015), (Lin et al., 2015)).”
Pi-Pi contacts are an overlooked protein feature relevant to phase separation (2018, background): “2011), FUS (Burke et al., 2015; Kato et al., 2012), Ddx4 and the homologous LAF-1 (Nott et al.”
Tau protein liquid–liquid phase separation can initiate tau aggregation (2018, neutral): “For example, FUS, TDP-43, and hnRNPA1 contain “prion-like” LCDs that drive their phase separation.”
Phase separation in biology; functional organization of a higher order (2016, background): “liquid-like droplets [42, 44, 46, 53, 55] recover after photobleaching on a timescale of seconds to tens of seconds.”
Phosphorylation of the FUS low‐complexity domain disrupts phase separation, aggregation, and toxicity (2017, neutral): “However, full-length FUS droplets appear much more viscous and heterogeneous than previously characterized FUS LC droplets (Burke et al , 2015), as suggested by incomplete recovery after photobleaching at time points as early as 45 min after TEV cleavage.”
RNA-Induced Conformational Switching and Clustering of G3BP Drive Stress Granule Assembly by Condensation (2020, background): “This was substantially lower than estimates for protein-only condensates, which can reach 400 mg/mL (Brady et al., 2017; Burke et al., 2015).”
A molecular grammar governing the driving forces for phase separation of prion-like RNA binding proteins (2018, neutral): “Our experiments reinforce previous in vitro observations that isolated PLDs of the FET proteins have very different phase behavior than the full-length proteins (Burke et al., 2015; Han et al., 2012; Lin et al., 2015).”
ALS mutations disrupt phase separation mediated by -helical structure in the TDP-43 low complexity C-terminal domain (2016, background): “TDP-43 C-terminal domain undergoes liquid-liquid phase separation Because the low complexity domain of FUS is sufficient for LLPS(Burke et al., 2015), we tested if the C-terminal domain of TDP-43 is similarly sufficient.”