FtsZ‐Induced Shape Transformation of Coacervates

Fanalista, Federico; Deshpande, Siddharth; Lau, Anson; Pawlik, Grzegorz; Dekker, Cees

doi:10.1002/adbi.201800136

Cited by 28 publications

(26 citation statements)

References 36 publications

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“…As a means to quantify the degree of protein localization in a host condensate, we measured the partition coefficient of the protein to be P FtsZ = 14.2 ± 3.8 ( n = 25, see Methods for details). The observed homogenous distribution contrasts the FtsZ localization at the interface of pLL/ATP coacervates that was reported recently 11 , where FtsZ was added to a solution containing pre-formed, stable coacervates. In the current case, however, FtsZ was already present when the phase separation occurred.…”

Section: Resultscontrasting

confidence: 79%

Spatiotemporal control of coacervate formation within liposomes

et al. 2019

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Liquid-liquid phase separation (LLPS), especially coacervation, plays a crucial role in cell biology, as it forms numerous membraneless organelles in cells. Coacervates play an indispensable role in regulating intracellular biochemistry, and their dysfunction is associated with several diseases. Understanding of the LLPS dynamics would greatly benefit from controlled in vitro assays that mimic cells. Here, we use a microfluidics-based methodology to form coacervates inside cell-sized (~10 µm) liposomes, allowing control over the dynamics. Protein-pore-mediated permeation of small molecules into liposomes triggers LLPS passively or via active mechanisms like enzymatic polymerization of nucleic acids. We demonstrate sequestration of proteins (FtsZ) and supramolecular assemblies (lipid vesicles), as well as the possibility to host metabolic reactions ( β - galactosidase activity) inside coacervates. This coacervate-in-liposome platform provides a versatile tool to understand intracellular phase behavior, and these hybrid systems will allow engineering complex pathways to reconstitute cellular functions and facilitate bottom-up creation of synthetic cells.

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Section: Resultscontrasting

confidence: 79%

Spatiotemporal control of coacervate formation within liposomes

et al. 2019

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“…Although scaffolds are the drivers of condensate formation, the solution conditions and concentration regimes in which formation and dissolution of condensates occurs can be regulated by controlling the concentrations of non-scaffold molecules (17,18) or via post-translational modifications to scaffolds (16,19). An example of regulation by non-scaffold molecules comes from recent work on stress granules showing the different effects of Caprin-1 and USP-10 on the stability and integrity of stress granules (12,20,21).…”

Section: Significancementioning

confidence: 99%

Ligand Effects on Phase Separation of Multivalent Macromolecules

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Dar

Pappu

2020

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Biomolecular condensates are thought to form via phase transitions of multivalent macromolecules. Components of condensates have been classified as scaffolds and clients. In this classification, scaffolds drive condensate formation whereas clients partition into condensates. However, non-scaffold molecules can be ligands that modulate the phase behavior of scaffolds via preferential binding across phase boundaries. Here, we present computational results that explain how preferential binding of ligands to scaffold molecules in different phases can impact phase separation and the compositions of dense phases. We show that phase separation is destabilized by monovalent ligands. In contrast, multivalent ligands, depending on their mode of binding, can stabilize phase separation by serving as crosslinkers that network scaffold molecules. We also find that concentrations of scaffolds within dense phases are generally diluted by ligands. This arises because ligands destabilize condensates by preferentially binding to scaffolds in the dilute phase or because ligands have to be accommodated within dense phases when they bind preferentially to scaffolds in the dense phase. Importantly, we show that partition coefficients, which are routinely used for compositional profiling of condensates, say nothing about the regulatory impact of ligands on the phase behavior of scaffolds. Therefore, instead of measuring partition coefficients it becomes imperative to measure how ligands impact threshold concentrations of scaffolds. These measurements, performed as a function of ligand concentration, will help with understanding the regulation of condensates and enable the design of molecules that impact condensate formation or dissolution.

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“…[151] Spontaneous sequestration of actin filaments in PEGsuspended dextran-rich microdroplets was also reported, and the accumulation of bundled filaments at the droplets urface was shown to induce dropletdeformations. [153] Shape instabilities and self-division were also reported in anisotropic liquid droplets formed by actin filaments. Complex polypeptide-based coacervates were shown to sequester actin spontaneously,f or instance, resulting in an enhanced actin filament assembly rate due to the localised enrichment in cytoskeletal proteins.…”

Section: Towards Droplet Division Through Cytoskeleton Reconstitutionmentioning

confidence: 91%

Dynamic Synthetic Cells Based on Liquid–Liquid Phase Separation

2019

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Living cells have long been a source of inspiration for chemists. Their capacity of performing complex tasks relies on the spatiotemporal coordination of matter and energy fluxes. Recent years have witnessed growing interest in the bottom‐up construction of cell‐like models capable of reproducing aspects of such dynamic organisation. Liquid–liquid phase‐separation (LLPS) processes in water are increasingly recognised as representing a viable compartmentalisation strategy through which to produce dynamic synthetic cells. Herein, we highlight examples of the dynamic properties of LLPS used to assemble synthetic cells, including their biocatalytic activity, reversible condensation and dissolution, growth and division, and recent directions towards the design of higher‐order structures and behaviour.

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FtsZ‐Induced Shape Transformation of Coacervates

Cited by 28 publications

References 36 publications

Spatiotemporal control of coacervate formation within liposomes

Spatiotemporal control of coacervate formation within liposomes

Ligand Effects on Phase Separation of Multivalent Macromolecules

Dynamic Synthetic Cells Based on Liquid–Liquid Phase Separation

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