Non-spherical coacervate shapes in an enzyme driven active system
Reads0
Chats0
TLDR
This work presents and characterize a system of enzymatically active coacervates containing spermine, RNA, free nucleotides, and the template independent RNA (de)polymerase PNPase, and finds that these RNA coACervates display transient non-spherical shapes, and systematically study how P NPase concentration, UDP concentration and temperature affect coacervation morphology.Abstract:
Coacervates are polymer-rich droplets that form through liquid-liquid phase separation in polymer solutions. Liquid-liquid phase separation and coacervation have recently been shown to play an important role in the organization of biological systems. Such systems are highly dynamic and under continuous influence of enzymatic and chemical processes. However, it is still unclear how enzymatic and chemical reactions affect the coacervation process. Here, we present and characterize a system of enzymatically active coacervates containing spermine, RNA, free nucleotides, and the template independent RNA (de)polymerase PNPase. We find that these RNA coacervates display transient non-spherical shapes, and we systematically study how PNPase concentration, UDP concentration and temperature affect coacervate morphology. Furthermore, we show that PNPase localizes predominantly into the coacervate phase and that its depolymerization activity in high-phosphate buffer causes coacervate degradation. Our observations of non-spherical coacervate shapes may have broader implications for the relationship between (bio-)chemical activity and coacervate biology.read more
References
More filters
Journal ArticleDOI
Phase Transition of a Disordered Nuage Protein Generates Environmentally Responsive Membraneless Organelles
Timothy J. Nott,Timothy J. Nott,Evangelia Petsalaki,Patrick Farber,Dylan Jervis,Eden Fussner,Anne Plochowietz,Timothy D. Craggs,David P. Bazett-Jones,Tony Pawson,Julie D. Forman-Kay,Andrew Baldwin +11 more
TL;DR: It is demonstrated that the disordered tails of Ddx4, a primary constituent of nuage or germ granules, form phase-separated organelles both in live cells and in vitro, and proposed that phase separation of disordered proteins containing weakly interacting blocks is a general mechanism for forming regulated, membraneless organlles.
Journal ArticleDOI
Polymer physics of intracellular phase transitions
TL;DR: These findings highlight the relevance of classical concepts from the physics of polymeric phase transitions for understanding the assembly of intracellular membrane-less compartments, and challenge the challenge of applying these concepts given the heteropolymeric nature of protein sequences, the complex intrACEllular environment, and non-equilibrium features intrinsic to cells.
Journal ArticleDOI
Active liquid-like behavior of nucleoli determines their size and shape in Xenopus laevis oocytes.
TL;DR: It is shown that both the size and shape of the amphibian oocyte nucleolus ultimately arise because nucleoli behave as liquid-like droplets of RNA and protein, exhibiting characteristic viscous fluid dynamics even on timescales of < 1 min.
Journal ArticleDOI
ALS/FTD Mutation-Induced Phase Transition of FUS Liquid Droplets and Reversible Hydrogels into Irreversible Hydrogels Impairs RNP Granule Function
Tetsuro Murakami,Seema Qamar,Julie Qiaojin Lin,Gabriele S. Kaminski Schierle,Eric J. Rees,Akinori Miyashita,Ana Rita Costa,Roger B. Dodd,Fiona T. S. Chan,Claire H. Michel,Deborah Kronenberg-Versteeg,Yi Li,Seung-Pil Yang,Yosuke Wakutani,William Meadows,Rodylyn Rose Ferry,Liang Dong,Gian Gaetano Tartaglia,Gian Gaetano Tartaglia,Giorgio Favrin,Wen-Lang Lin,Dennis W. Dickson,Mei Zhen,David Ron,Gerold Schmitt-Ulms,Paul E. Fraser,Neil A. Shneider,Christine E. Holt,Michele Vendruscolo,Clemens F. Kaminski,Peter St George-Hyslop,Peter St George-Hyslop +31 more
TL;DR: A model in which low-complexity (LC) domains of FUS drive its physiologically reversible assembly into membrane-free, liquid droplet and hydrogel-like structures mitigates neurotoxicity is proposed and suggests a potential therapeutic strategy that may also be applicable to ALS/FTD associated with mutations in other RNA binding proteins.
Journal ArticleDOI
Liquid–liquid phase separation of the microtubule-binding repeats of the Alzheimer-related protein Tau
Susmitha Ambadipudi,Jacek Biernat,Dietmar Riedel,Eckhard Mandelkow,Markus Zweckstetter,Markus Zweckstetter +5 more
TL;DR: The authors identify conditions, where the microtubule-binding repeats of Tau undergo a phosphorylation-dependent liquid–liquid phase separation, leading to molecular crowding in the formed Tau liquid droplets and characterize them by NMR and other biophysical methods.