Hallegger_Chakrabarti_Lee_TDP-43 condensation properties_Mendeley
Mutations causing amyotrophic lateral sclerosis (ALS) often affect the condensation properties of RNA-binding proteins (RBPs). However, the role of RBP condensation in the specificity and functions of protein-RNA complexes remains unclear. We created a series of TDP-43 C-terminal domain (CTD) variants, which exhibited a gradient of low to high condensation propensity, as observed in vitro and by nuclear mobility and foci formation. Notably, capacity for condensation was required for efficient TDP-43 assembly on subsets of RNA-binding regions, which contain unusually long clusters of motifs of characteristic types and density. These ‘binding-region condensates’ are promoted by homomeric CTD-driven interactions and are required for efficient regulation of a subset of bound transcripts, including autoregulation of TDP-43 mRNA. Thus, we establish that RBP condensation can occur in a binding-region specific manner to selectively modulate transcriptome-wide RNA regulation, which has implications for remodeling RNA networks in the context of signaling, disease, and evolution.
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Data linked to METHODS In vitro phase separation assays Microscopy and imaging analysis Fluorescence recovery after photobleaching (FRAP) Comparison of in vitro Csat with FRAP and foci measurements Western blot analysis RT-PCR