The post-translational modification SUMO affects TDP-43 phase separation, compartmentalization, and aggregation in a zebrafish model

Maurel, Cindy; Scherer, Natalie M.; Hogan, Alison; Vidal-Itriago, Andrés; Don, Emily K.; Radford, Rowan A. W.; Chapman, Tyler; Cull, Stephen; Vourc’h, Patrick; Chung, Roger S.; Lee, Albert; Morsch, Marco

doi:10.1101/2022.08.14.503569

Cited by 2 publications

(2 citation statements)

References 91 publications

(133 reference statements)

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“…The functional effects of the G294V mutation are yet to be fully established. Our zebrafish observations did not display significantly altered compartment specific expression of hTDP-43 G294V compared to hTDP-43 WT (this study and ( 75 )). These findings align with two other studies reporting unaltered cytoplasmic expression levels in primary patient fibroblasts ( 74 ) and iPSCs ( 76 ).…”

Section: Discussionsupporting

confidence: 53%

RNA-binding properties orchestrate TDP-43 homeostasis through condensate formation in vivo

Scherer,

Maurel,

Graus

et al. 2024

Nucleic Acids Research

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Insoluble cytoplasmic aggregate formation of the RNA-binding protein TDP-43 is a major hallmark of neurodegenerative diseases including Amyotrophic Lateral Sclerosis. TDP-43 localizes predominantly in the nucleus, arranging itself into dynamic condensates through liquid–liquid phase separation (LLPS). Mutations and post-translational modifications can alter the condensation properties of TDP-43, contributing to the transition of liquid-like biomolecular condensates into solid-like aggregates. However, to date it has been a challenge to study the dynamics of this process in vivo. We demonstrate through live imaging that human TDP-43 undergoes nuclear condensation in spinal motor neurons in a living animal. RNA-binding deficiencies as well as post-translational modifications can lead to aberrant condensation and altered TDP-43 compartmentalization. Single-molecule tracking revealed an altered mobility profile for RNA-binding deficient TDP-43. Overall, these results provide a critically needed in vivo characterization of TDP-43 condensation, demonstrate phase separation as an important regulatory mechanism of TDP-43 accessibility, and identify a molecular mechanism of how functional TDP-43 can be regulated.

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Section: Discussionsupporting

confidence: 53%

RNA-binding properties orchestrate TDP-43 homeostasis through condensate formation in vivo

Scherer,

Maurel,

Graus

et al. 2024

Nucleic Acids Research

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“…Our tool quantifies condensate size and concentration to obtain detailed information on the steps from initial formation of small nuclei to nanocondensates then on to micron sized droplets. As monitoring protein LLPS occurring in cells is becoming an active field of research [40][41][42][43][44] , we envision our method as a useful tool to support findings of nanocondensation in cells 45,46 . Our assay can be applied to different proteins and conditions ranging from buffers to crowded cellular environments.…”

Section: Discussionmentioning

confidence: 97%

Quantification of nanocondensates formation at the single molecule level

Houx,

Copie,

Gambin

et al. 2024

Preprint

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Understanding the molecular mechanisms of biomolecular condensate formation through liquid-liquid phase separation is crucial for deciphering cellular cues in normal and pathological contexts. Recent studies have highlighted the existence of sub-micron assemblies, known as nanocondensates or mesoscopic clusters, in the organization of a significant portion of the proteome. However, as smaller condensates are invisible to classical microscopy, new tools must be developed to quantify their numbers and properties. Here, we establish a simple analysis framework using single molecule fluorescence spectroscopy to quantify the formation of nanocondensates diffusing in solution. We used the low-complexity domain of TAR DNA-binding protein 43 (TDP-43) as a model system to show that we can recapitulate the phase separation diagram of the protein in various conditions. Single molecule spectroscopy reveals rapid formation of TDP-43 nanoclusters at ten-fold lower concentrations than described previously by microscopy. We demonstrate how straightforward fingerprinting of individual nanocondensates provides an exquisite quantification of their formation, size, density, and their temporal evolution. Overall, this study highlights the potential of single molecule spectroscopy to investigate the formation of biomolecular condensates and liquid-liquid phase separation mechanisms in protein systems.

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The post-translational modification SUMO affects TDP-43 phase separation, compartmentalization, and aggregation in a zebrafish model

Cited by 2 publications

References 91 publications

RNA-binding properties orchestrate TDP-43 homeostasis through condensate formation in vivo

RNA-binding properties orchestrate TDP-43 homeostasis through condensate formation in vivo

Quantification of nanocondensates formation at the single molecule level

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