| Publication note | By combining NMR and Raman spectroscopy, mutagenesis, and molecular simulation, we demonstrate that heterogeneous interactions involving all residue types underlie LLPS of human FUS LC. We find no evidence that FUS LC adopts conformations with traditional secondary structure elements in the condensed phase, rather it maintains conformational heterogeneity. We show that hydrogen bonding, π/sp2 and hydrophobic interactions all contribute to stabilizing LLPS of FUS LC. In addition to contributions from tyrosine residues, we find that glutamine residues participate in contacts leading to LLPS of FUS LC. (Organism: Homo sapiens; Cell line: _) |
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| Material state | liquid : We demonstrate that the condensed phase is devoid of detectable static structures and that amorphous aggregates of FUS LC are structurally different from the liquid phase. | ||
| LLPS region | IDR region | Key domains | low-complexity domain(LCD) : FUS LC LLPS is governed by non-specific hydrogen bonding, hydrophobic and π/sp2 interactions that involve all of the residue types within the sequence. |
| Protein partner | _ ( _ ) |
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|---|---|---|
| RNA partner | _ |
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| Others | _ |
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| PTM | _ |
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| Mutation | p.Q27S/Q28S/Q93S/Q94S/Q132S/Q133S/Q157S/Q158S,p.Q8S/Q9S/Q60S/Q62S/Q102S/Q103S/Q139S/Q140S,p.S30Q/S44Q/S48Q/S53Q/S70Q/S84Q/S89Q/S95Q/S108Q/S117Q/S119Q/S127Q,p.Y6F/Y14F/Y17F/Y25F/Y33F/Y38F/Y41F/Y50F/Y55F/Y58F/Y66F/Y75F/Y81F/Y91F/Y97F/Y100F/Y113F/Y122F/Y130F/Y136F/Y143F/Y149F/Y155F/Y161F |
Reduced glutamine content (p.Q27S/Q28S/Q93S/Q94S/Q132S/Q133S/Q157S/Q158S, p.Q8S/Q9S/Q60S/Q62S/Q102S/Q103S/Q139S/Q140S) resulted in higher concentration of protein remaining in the supernatant compared to WT, suggesting that removal of glutamine reduced phase separation. Increased glutamine content (p.S30Q/S44Q/S48Q/S53Q/S70Q/S84Q/S89Q/S95Q/S108Q/S117Q/S119Q/S127Q) decreased protein concentration in the supernatant compared to the WT, suggesting that increasing glutamine content promotes phase separation; We did attempt to evaluate the effect of increasing hydrophobicity by changing all twenty-four FUS LC tyrosines to phenylalanine(p.Y6F/Y14F/Y17F/Y25F/Y33F/Y38F/Y41F/Y50F/Y55F/Y58F/Y66F/Y75F/Y81F/Y91F/Y97F/Y100F/Y113F/Y122F/Y130F/Y136F/Y143F/Y149F/Y155F/Y161F). However, this resulted in a dramatically more aggregation-prone FUS LC and full-length FUS. |
| Alternative splicing | _ | |
| Repeat | _ | |
| Oligomerization | _ | |