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TDP-43 ?-helical structure tunes liquid-liquid phase separation and function.


ABSTRACT: Liquid-liquid phase separation (LLPS) is involved in the formation of membraneless organelles (MLOs) associated with RNA processing. The RNA-binding protein TDP-43 is present in several MLOs, undergoes LLPS, and has been linked to the pathogenesis of amyotrophic lateral sclerosis (ALS). While some ALS-associated mutations in TDP-43 disrupt self-interaction and function, here we show that designed single mutations can enhance TDP-43 assembly and function via modulating helical structure. Using molecular simulation and NMR spectroscopy, we observe large structural changes upon dimerization of TDP-43. Two conserved glycine residues (G335 and G338) are potent inhibitors of helical extension and helix-helix interaction, which are removed in part by variants at these positions, including the ALS-associated G335D. Substitution to helix-enhancing alanine at either of these positions dramatically enhances phase separation in vitro and decreases fluidity of phase-separated TDP-43 reporter compartments in cells. Furthermore, G335A increases TDP-43 splicing function in a minigene assay. Therefore, the TDP-43 helical region serves as a short but uniquely tunable module where application of biophysical principles can precisely control assembly and function in cellular and synthetic biology applications of LLPS.

SUBMITTER: Conicella AE 

PROVIDER: S-EPMC7084079 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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TDP-43 α-helical structure tunes liquid-liquid phase separation and function.

Conicella Alexander E AE   Dignon Gregory L GL   Zerze Gül H GH   Schmidt Hermann Broder HB   D'Ordine Alexandra M AM   Kim Young C YC   Rohatgi Rajat R   Ayala Yuna M YM   Mittal Jeetain J   Fawzi Nicolas L NL  

Proceedings of the National Academy of Sciences of the United States of America 20200304 11


Liquid-liquid phase separation (LLPS) is involved in the formation of membraneless organelles (MLOs) associated with RNA processing. The RNA-binding protein TDP-43 is present in several MLOs, undergoes LLPS, and has been linked to the pathogenesis of amyotrophic lateral sclerosis (ALS). While some ALS-associated mutations in TDP-43 disrupt self-interaction and function, here we show that designed single mutations can enhance TDP-43 assembly and function via modulating helical structure. Using mo  ...[more]

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