Liquid-liquid Phase Separation Induces Pathogenic Tau Conformations in Vitro

Overview

Journal Nat Commun

Specialty Biology

Date 2020 Jun 6

PMID 32499559

Citations 126

Authors

Nicholas M Kanaan

Chelsey Hamel

Tessa Grabinski

Benjamin Combs

Affiliations

Soon will be listed here.

Abstract

Formation of membrane-less organelles via liquid-liquid phase separation is one way cells meet the biological requirement for spatiotemporal regulation of cellular components and reactions. Recently, tau, a protein known for its involvement in Alzheimer's disease and other tauopathies, was found to undergo liquid-liquid phase separation making it one of several proteins associated with neurodegenerative diseases to do so. Here, we demonstrate that tau forms dynamic liquid droplets in vitro at physiological protein levels upon molecular crowding in buffers that resemble physiological conditions. Tau droplet formation is significantly enhanced by disease-associated modifications, including the AT8 phospho-epitope and the P301L tau mutation linked to an inherited tauopathy. Moreover, tau droplet dynamics are significantly reduced by these modified forms of tau. Extended phase separation promoted a time-dependent adoption of toxic conformations and oligomerization, but not filamentous aggregation. P301L tau protein showed the greatest oligomer formation following extended phase separation. These findings suggest that phase separation of tau may facilitate the formation of non-filamentous pathogenic tau conformations.

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References

Brangwynne C . Phase transitions and size scaling of membrane-less organelles. J Cell Biol. 2013; 203(6):875-81. PMC: 3871435. DOI: 10.1083/jcb.201308087. View

Hyman A, Weber C, Julicher F . Liquid-liquid phase separation in biology. Annu Rev Cell Dev Biol. 2014; 30:39-58. DOI: 10.1146/annurev-cellbio-100913-013325. View

Handwerger K, Cordero J, Gall J . Cajal bodies, nucleoli, and speckles in the Xenopus oocyte nucleus have a low-density, sponge-like structure. Mol Biol Cell. 2004; 16(1):202-11. PMC: 539164. DOI: 10.1091/mbc.e04-08-0742. View

Dundr M, Misteli T . Biogenesis of nuclear bodies. Cold Spring Harb Perspect Biol. 2010; 2(12):a000711. PMC: 2982170. DOI: 10.1101/cshperspect.a000711. View

Souquere S, Mollet S, Kress M, Dautry F, Pierron G, Weil D . Unravelling the ultrastructure of stress granules and associated P-bodies in human cells. J Cell Sci. 2009; 122(Pt 20):3619-26. DOI: 10.1242/jcs.054437. View

Yang Z, Jakymiw A, Wood M, Eystathioy T, Rubin R, Fritzler M . GW182 is critical for the stability of GW bodies expressed during the cell cycle and cell proliferation. J Cell Sci. 2004; 117(Pt 23):5567-78. DOI: 10.1242/jcs.01477. View

Pak C, Kosno M, Holehouse A, Padrick S, Mittal A, Ali R . Sequence Determinants of Intracellular Phase Separation by Complex Coacervation of a Disordered Protein. Mol Cell. 2016; 63(1):72-85. PMC: 4973464. DOI: 10.1016/j.molcel.2016.05.042. View

Lin Y, Protter D, Rosen M, Parker R . Formation and Maturation of Phase-Separated Liquid Droplets by RNA-Binding Proteins. Mol Cell. 2015; 60(2):208-19. PMC: 4609299. DOI: 10.1016/j.molcel.2015.08.018. View

Aumiller Jr W, Keating C . Experimental models for dynamic compartmentalization of biomolecules in liquid organelles: Reversible formation and partitioning in aqueous biphasic systems. Adv Colloid Interface Sci. 2016; 239:75-87. DOI: 10.1016/j.cis.2016.06.011. View

10.

Wright P, Dyson H . Intrinsically disordered proteins in cellular signalling and regulation. Nat Rev Mol Cell Biol. 2014; 16(1):18-29. PMC: 4405151. DOI: 10.1038/nrm3920. View

11.

Avila J, Jimenez J, Sayas C, Bolos M, Zabala J, Rivas G . Tau Structures. Front Aging Neurosci. 2016; 8:262. PMC: 5099159. DOI: 10.3389/fnagi.2016.00262. View

12.

Sjoberg M, Shestakova E, Mansuroglu Z, Maccioni R, Bonnefoy E . Tau protein binds to pericentromeric DNA: a putative role for nuclear tau in nucleolar organization. J Cell Sci. 2006; 119(Pt 10):2025-34. DOI: 10.1242/jcs.02907. View

13.

Brunello C, Yan X, Huttunen H . Internalized Tau sensitizes cells to stress by promoting formation and stability of stress granules. Sci Rep. 2016; 6:30498. PMC: 4962319. DOI: 10.1038/srep30498. View

14.

Vanderweyde T, Apicco D, Youmans-Kidder K, Ash P, Cook C, Lummertz da Rocha E . Interaction of tau with the RNA-Binding Protein TIA1 Regulates tau Pathophysiology and Toxicity. Cell Rep. 2016; 15(7):1455-1466. PMC: 5325702. DOI: 10.1016/j.celrep.2016.04.045. View

15.

Hernandez-Vega A, Braun M, Scharrel L, Jahnel M, Wegmann S, Hyman B . Local Nucleation of Microtubule Bundles through Tubulin Concentration into a Condensed Tau Phase. Cell Rep. 2017; 20(10):2304-2312. PMC: 5828996. DOI: 10.1016/j.celrep.2017.08.042. View

16.

Zhang X, Lin Y, Eschmann N, Zhou H, Rauch J, Hernandez I . RNA stores tau reversibly in complex coacervates. PLoS Biol. 2017; 15(7):e2002183. PMC: 5500003. DOI: 10.1371/journal.pbio.2002183. View

17.

Lin Y, Fichou Y, Zeng Z, Hu N, Han S . Electrostatically Driven Complex Coacervation and Amyloid Aggregation of Tau Are Independent Processes with Overlapping Conditions. ACS Chem Neurosci. 2020; 11(4):615-627. DOI: 10.1021/acschemneuro.9b00627. View

18.

Majumdar A, Dogra P, Maity S, Mukhopadhyay S . Liquid-Liquid Phase Separation Is Driven by Large-Scale Conformational Unwinding and Fluctuations of Intrinsically Disordered Protein Molecules. J Phys Chem Lett. 2019; 10(14):3929-3936. DOI: 10.1021/acs.jpclett.9b01731. View

19.

Ukmar-Godec T, Hutten S, Grieshop M, Rezaei-Ghaleh N, Cima-Omori M, Biernat J . Lysine/RNA-interactions drive and regulate biomolecular condensation. Nat Commun. 2019; 10(1):2909. PMC: 6606616. DOI: 10.1038/s41467-019-10792-y. View

20.

Ambadipudi S, Reddy J, Biernat J, Mandelkow E, Zweckstetter M . Residue-specific identification of phase separation hot spots of Alzheimer's-related protein tau. Chem Sci. 2019; 10(26):6503-6507. PMC: 6610569. DOI: 10.1039/c9sc00531e. View