| Publication note | Exposure of cell or tissue lysates to a biotinylated isoxazole (b-isox) chemical precipitated hundreds of RNA binding proteins with significant overlap to the constituents of RNA granules. The LC sequences within these proteins are both necessary and sufficient for b-isox-mediated aggregation, and these domains can undergo a concentration-dependent phase transition to a hydrogel-like state in the absence of the chemical; Homotypic trapping of the GST:FUS LC by the mCherry:FUS LC hydrogel was sufficiently avid that GFP signal hardly decayed over the hour-long washout period. Hydrogel droplets of the mCherry:FUS LC domain of roughly 500um in diameter (Organism: Homo sapiens; Cell line: _) |
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| Material state | hydrogel : The LC sequences within these proteins are both necessary and sufficient for b-isox-mediated aggregation, and these domains can undergo a concentration-dependent phase transition to a hydrogel-like state in the absence of the chemical. | ||
| LLPS region | 2-214 | Key domains | low-complexity domain(LCD) : The LC domains are both necessary and sufficient for b-isox-mediated precipitation, for hydrogel formation, and for retention by pre-formed hydrogel droplet. |
| Protein partner | hnRNPA2,RBM3,hnRNPA1,TIA1,CPEB2,FMRP,CIRBP,TDP43,Sup35 ( P22626,P98179,P09651,P31483,Q7Z5Q1,Q06787,Q14011,Q13148,P05453 ) |
Homotypic trapping of the GST:FUS LC by the mCherry:FUS LC hydrogel was sufficiently avid that GFP signal hardly decayed over the hour-long washout period. The LC domains of hnRNPA2 and RBM3 RNA binding proteins adhered to the hydrogel with moderate apparent avidity. The hnRNPA1, TIA1 and CPEB2 LC domains bound to the hydrogel with more modest avidity. Binding of the LC domains of four additional test samples (FMRP, CIRBP, TDP43 and yeast Sup35) was also observed |
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| RNA partner | _ |
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| Others | _ |
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| PTM | _ |
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| Mutation | p.Y17S/Y75S/Y81S/Y143S/Y149S,p.Y14S/Y17S/Y33S/Y38S/Y41S/Y130S/Y143S/Y149S/Y155S,p.Y14S/Y17S/Y33S/Y38S/Y41S/Y55S/Y58S/Y91S/Y97S/Y100S/Y130S/Y143S/Y149S/Y155S/Y161S,p.Y6S/Y14S/Y17S/Y25S/Y33S/Y38S/Y41S/Y50S/Y55S/Y58S/Y66S/Y75S/Y81S/Y91S/Y97S/Y100S/Y113S/Y122S/Y130S/Y136S/Y143S/Y149S/Y155S/Y161S/Y177S/Y194S/Y208S |
we prepared mutated derivatives of GFP:FUS LC wherein varying numbers of tyrosine residues in the [G/S]Y[G/S] motifs were mutated to serine. The S1 mutant(p.Y17S/Y75S/Y81S/Y143S/Y149S) changed 5 of the relevant tyrosine residues in the LC domain of FUS to serine. The S2(p.Y14S/Y17S/Y33S/Y38S/Y41S/Y130S/Y143S/Y149S/Y155S) and S3(p.Y14S/Y17S/Y33S/Y38S/Y41S/Y55S/Y58S/Y91S/Y97S/Y100S/Y130S/Y143S/Y149S/Y155S/Y161S) variants mutated 9 and 15 of the tyrosines to serine respectively, and the allS variant mutated all 27 tyrosines to serine. Upon purification and concentration, none of the mutants were capable of hydrogel formation. When these proteins were assayed for binding to mCherry:FUS LC hydrogel, moderate retention was observed for the S1 mutant, weaker retention was observed for S2, and little or no retention was observed for S3 and allS(p.Y6S/Y14S/Y17S/Y25S/Y33S/Y38S/Y41S/Y50S/Y55S/Y58S/Y66S/Y75S/Y81S/Y91S/Y97S/Y100S/Y113S/Y122S/Y130S/Y136S/Y143S/Y149S/Y155S/Y161S/Y177S/Y194S/Y208S). |
| Alternative splicing | _ | |
| Repeat | _ | |
| Oligomerization | _ | |