Sequence determinants of protein phase behavior from a coarse-grained model

Dignon, Gregory L.; Zheng, Wenwei; Kim, Young C.; Best, Robert B.; Mittal, Jeetain; Ofran, Yanay

doi:10.1371/journal.pcbi.1005941

Title: Sequence determinants of protein phase behavior from a coarse-grained model

Journal Article · 24 January 2018 · PLoS Computational Biology (Online)

DOI: https://doi.org/10.1371/journal.pcbi.1005941 · OSTI ID:1419611

^[1]; Zheng, Wenwei ^[2]; Kim, Young C. ^[3]; Best, Robert B. ^[4];

^[1]; Ofran, Yanay ^[5]

Lehigh Univ., Bethlehem, PA (United States)
National Inst. of Health (NIH), Bethesda, MD (United States); ; Arizona State Univ., Mesa, AZ (United States)
Naval Research Lab. (NRL), Washington, DC (United States)
National Inst. of Health (NIH), Bethesda, MD (United States)
Bar Ilan Univ., Ramat Gan, Tel Aviv (Israel)

Membraneless organelles important to intracellular compartmentalization have recently been shown to comprise assemblies of proteins which undergo liquid-liquid phase separation (LLPS). However, many proteins involved in this phase separation are at least partially disordered. The molecular mechanism and the sequence determinants of this process are challenging to determine experimentally owing to the disordered nature of the assemblies, motivating the use of theoretical and simulation methods. This work advances a computational framework for conducting simulations of LLPS with residue-level detail, and allows for the determination of phase diagrams and coexistence densities of proteins in the two phases. The model includes a short-range contact potential as well as a simplified treatment of electrostatic energy. Interaction parameters are optimized against experimentally determined radius of gyration data for multiple unfolded or intrinsically disordered proteins (IDPs). These models are applied to two systems which undergo LLPS: the low complexity domain of the RNA-binding protein FUS and the DEAD-box helicase protein LAF-1. We develop a novel simulation method to determine thermodynamic phase diagrams as a function of the total protein concentration and temperature. We show that the model is capable of capturing qualitative changes in the phase diagram due to phosphomimetic mutations of FUS and to the presence or absence of the large folded domain in LAF-1. We also explore the effects of chain-length, or multivalency, on the phase diagram, and obtain results consistent with Flory-Huggins theory for polymers. Moreover, the methodology presented here is flexible so that it can be easily extended to other pair potentials, be used with other enhanced sampling methods, and may incorporate additional features for biological systems of interest.

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Research Organization:: Lehigh Univ., Bethlehem, PA (United States); University of California, Berkeley, CA (United States)

Sponsoring Organization:: USDOE

Grant/Contract Number:: SC0013979; AC02-05CH11231

OSTI ID:: 1419611

Alternate ID(s):: OSTI ID: 1417943; OSTI ID: 1499874

Journal Information:: PLoS Computational Biology (Online), Vol. 14, Issue 1; ISSN 1553-7358

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 268 works

Citation information provided by
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References (140)

RNA Remodeling Activity of DEAD Box Proteins Tuned by Protein Concentration, RNA Length, and ATP Kim, Younghoon; Myong, Sua Molecular Cell, Vol. 63, Issue 5 https://doi.org/10.1016/j.molcel.2016.07.010	journal	September 2016
Phase Transition of Spindle-Associated Protein Regulate Spindle Apparatus Assembly Jiang, Hao; Wang, Shusheng; Huang, Yuejia Cell, Vol. 163, Issue 1 https://doi.org/10.1016/j.cell.2015.08.010	journal	September 2015
Innovative scattering analysis shows that hydrophobic disordered proteins are expanded in water Riback, Joshua A.; Bowman, Micayla A.; Zmyslowski, Adam M. Science, Vol. 358, Issue 6360 https://doi.org/10.1126/science.aan5774	journal	October 2017
Consistent View of Polypeptide Chain Expansion in Chemical Denaturants from Multiple Experimental Methods Borgia, Alessandro; Zheng, Wenwei; Buholzer, Karin Journal of the American Chemical Society, Vol. 138, Issue 36 https://doi.org/10.1021/jacs.6b05917	journal	September 2016
Role of Repulsive Forces in Determining the Equilibrium Structure of Simple Liquids Weeks, John D.; Chandler, David; Andersen, Hans C. The Journal of Chemical Physics, Vol. 54, Issue 12 https://doi.org/10.1063/1.1674820	journal	June 1971
RNA Controls PolyQ Protein Phase Transitions Zhang, Huaiying; Elbaum-Garfinkle, Shana; Langdon, Erin M. Molecular Cell, Vol. 60, Issue 2 https://doi.org/10.1016/j.molcel.2015.09.017	journal	October 2015
How Fast-Folding Proteins Fold Lindorff-Larsen, K.; Piana, S.; Dror, R. O. Science, Vol. 334, Issue 6055 https://doi.org/10.1126/science.1208351	journal	October 2011
How accurate are polymer models in the analysis of Förster resonance energy transfer experiments on proteins? O’Brien, Edward P.; Morrison, Greg; Brooks, Bernard R. The Journal of Chemical Physics, Vol. 130, Issue 12 https://doi.org/10.1063/1.3082151	journal	March 2009
Phase behaviour of disordered proteins underlying low density and high permeability of liquid organelles Wei, Ming-Tzo; Elbaum-Garfinkle, Shana; Holehouse, Alex S. Nature Chemistry, Vol. 9, Issue 11 https://doi.org/10.1038/nchem.2803	journal	June 2017
RNA transcription modulates phase transition-driven nuclear body assembly Berry, Joel; Weber, Stephanie C.; Vaidya, Nilesh Proceedings of the National Academy of Sciences, Vol. 112, Issue 38 https://doi.org/10.1073/pnas.1509317112	journal	September 2015
ALS Mutations Disrupt Phase Separation Mediated by α-Helical Structure in the TDP-43 Low-Complexity C-Terminal Domain Conicella, Alexander E.; Zerze, Gül H.; Mittal, Jeetain Structure, Vol. 24, Issue 9 https://doi.org/10.1016/j.str.2016.07.007	journal	September 2016
Active liquid-like behavior of nucleoli determines their size and shape in Xenopus laevis oocytes Brangwynne, C. P.; Mitchison, T. J.; Hyman, A. A. Proceedings of the National Academy of Sciences, Vol. 108, Issue 11 https://doi.org/10.1073/pnas.1017150108	journal	February 2011
Vapor-liquid interfacial properties of fully flexible Lennard-Jones chains Blas, Felipe J.; MacDowell, Luis G.; de Miguel, Enrique The Journal of Chemical Physics, Vol. 129, Issue 14 https://doi.org/10.1063/1.2989115	journal	October 2008
Structural and hydrodynamic properties of an intrinsically disordered region of a germ cell-specific protein on phase separation Brady, Jacob P.; Farber, Patrick J.; Sekhar, Ashok Proceedings of the National Academy of Sciences, Vol. 114, Issue 39 https://doi.org/10.1073/pnas.1706197114	journal	September 2017
Mutations in the FUS/TLS Gene on Chromosome 16 Cause Familial Amyotrophic Lateral Sclerosis Kwiatkowski, T. J.; Bosco, D. A.; LeClerc, A. L. Science, Vol. 323, Issue 5918 https://doi.org/10.1126/science.1166066	journal	February 2009
Vapour–liquid phase equilibrium and surface tension of fully flexible Lennard–Jones chains Silmore, Kevin S.; Howard, Michael P.; Panagiotopoulos, Athanassios Z. Molecular Physics, Vol. 115, Issue 3 https://doi.org/10.1080/00268976.2016.1262075	journal	December 2016
Sequence-Specific Polyampholyte Phase Separation in Membraneless Organelles Lin, Yi-Hsuan; Forman-Kay, Julie D.; Chan, Hue Sun Physical Review Letters, Vol. 117, Issue 17 https://doi.org/10.1103/PhysRevLett.117.178101	journal	October 2016
Decoupling of size and shape fluctuations in heteropolymeric sequences reconciles discrepancies in SAXS vs. FRET measurements Fuertes, Gustavo; Banterle, Niccolò; Ruff, Kiersten M. Proceedings of the National Academy of Sciences, Vol. 114, Issue 31 https://doi.org/10.1073/pnas.1704692114	journal	July 2017
Net charge per residue modulates conformational ensembles of intrinsically disordered proteins Mao, A. H.; Crick, S. L.; Vitalis, A. Proceedings of the National Academy of Sciences, Vol. 107, Issue 18 https://doi.org/10.1073/pnas.0911107107	journal	April 2010
A Phase Separation Model for Transcriptional Control Hnisz, Denes; Shrinivas, Krishna; Young, Richard A. Cell, Vol. 169, Issue 1 https://doi.org/10.1016/j.cell.2017.02.007	journal	March 2017
Liquid demixing of intrinsically disordered proteins is seeded by poly(ADP-ribose) Altmeyer, Matthias; Neelsen, Kai J.; Teloni, Federico Nature Communications, Vol. 6, Issue 1 https://doi.org/10.1038/ncomms9088	journal	August 2015
Comparative Protein Modelling by Satisfaction of Spatial Restraints Šali, Andrej; Blundell, Tom L. Journal of Molecular Biology, Vol. 234, Issue 3 https://doi.org/10.1006/jmbi.1993.1626	journal	December 1993
Thermodynamics of High Polymer Solutions Flory, Paul J. The Journal of Chemical Physics, Vol. 10, Issue 1 https://doi.org/10.1063/1.1723621	journal	January 1942
The LC Domain of hnRNPA2 Adopts Similar Conformations in Hydrogel Polymers, Liquid-like Droplets, and Nuclei Xiang, Siheng; Kato, Masato; Wu, Leeju C. Cell, Vol. 163, Issue 4 https://doi.org/10.1016/j.cell.2015.10.040	journal	November 2015
Germline P Granules Are Liquid Droplets That Localize by Controlled Dissolution/Condensation Brangwynne, C. P.; Eckmann, C. R.; Courson, D. S. Science, Vol. 324, Issue 5935 https://doi.org/10.1126/science.1172046	journal	May 2009
Intrinsically disordered proteins as crucial constituents of cellular aqueous two phase systems and coacervates Uversky, Vladimir N.; Kuznetsova, Irina M.; Turoverov, Konstantin K. FEBS Letters, Vol. 589, Issue 1 https://doi.org/10.1016/j.febslet.2014.11.028	journal	November 2014
Some Properties of Solutions of Long-chain Compounds. Huggins, Maurice L. The Journal of Physical Chemistry, Vol. 46, Issue 1 https://doi.org/10.1021/j150415a018	journal	January 1942
A Simple Atomic-Level Hydrophobicity Scale Reveals Protein Interfacial Structure Kapcha, Lauren H.; Rossky, Peter J. Journal of Molecular Biology, Vol. 426, Issue 2 https://doi.org/10.1016/j.jmb.2013.09.039	journal	January 2014
Polymorphic transitions in single crystals: A new molecular dynamics method Parrinello, M.; Rahman, A. Journal of Applied Physics, Vol. 52, Issue 12 https://doi.org/10.1063/1.328693	journal	December 1981
Phase separation of signaling molecules promotes T cell receptor signal transduction Su, Xiaolei; Ditlev, Jonathon A.; Hui, Enfu Science, Vol. 352, Issue 6285 https://doi.org/10.1126/science.aad9964	journal	April 2016
The disordered P granule protein LAF-1 drives phase separation into droplets with tunable viscosity and dynamics Elbaum-Garfinkle, Shana; Kim, Younghoon; Szczepaniak, Krzysztof Proceedings of the National Academy of Sciences, Vol. 112, Issue 23 https://doi.org/10.1073/pnas.1504822112	journal	May 2015
In Vitro Reconstitution of a Cellular Phase-Transition Process that Involves the mRNA Decapping Machinery Fromm, Simon A.; Kamenz, Julia; Nöldeke, Erik R. Angewandte Chemie International Edition, Vol. 53, Issue 28 https://doi.org/10.1002/anie.201402885	journal	May 2014
Residue-by-Residue View of In Vitro FUS Granules that Bind the C-Terminal Domain of RNA Polymerase II Burke, Kathleen A.; Janke, Abigail M.; Rhine, Christy L. Molecular Cell, Vol. 60, Issue 2 https://doi.org/10.1016/j.molcel.2015.09.006	journal	October 2015
Stress-Triggered Phase Separation Is an Adaptive, Evolutionarily Tuned Response Riback, Joshua A.; Katanski, Christopher D.; Kear-Scott, Jamie L. Cell, Vol. 168, Issue 6 https://doi.org/10.1016/j.cell.2017.02.027	journal	March 2017
Water Dispersion Interactions Strongly Influence Simulated Structural Properties of Disordered Protein States Piana, Stefano; Donchev, Alexander G.; Robustelli, Paul The Journal of Physical Chemistry B, Vol. 119, Issue 16 https://doi.org/10.1021/jp508971m	journal	April 2015
Phase Separation by Low Complexity Domains Promotes Stress Granule Assembly and Drives Pathological Fibrillization Molliex, Amandine; Temirov, Jamshid; Lee, Jihun Cell, Vol. 163, Issue 1 https://doi.org/10.1016/j.cell.2015.09.015	journal	September 2015
The salting-out behavior of amides and its relation to the denaturation of proteins by salts Schrier, Eugene E.; Schrier, Evelyn B. The Journal of Physical Chemistry, Vol. 71, Issue 6 https://doi.org/10.1021/j100865a049	journal	May 1967
Formation and Maturation of Phase-Separated Liquid Droplets by RNA-Binding Proteins Lin, Yuan; Protter, David S. W.; Rosen, Michael K. Molecular Cell, Vol. 60, Issue 2 https://doi.org/10.1016/j.molcel.2015.08.018	journal	October 2015
Why are ?natively unfolded? proteins unstructured under physiologic conditions? Uversky, Vladimir N.; Gillespie, Joel R.; Fink, Anthony L. Proteins: Structure, Function, and Genetics, Vol. 41, Issue 3 https://doi.org/10.1002/1097-0134(20001115)41:3<415::AID-PROT130>3.0.CO;2-7	journal	January 2000
Protein crystallization and phase diagrams Asherie, N. Methods, Vol. 34, Issue 3 https://doi.org/10.1016/j.ymeth.2004.03.028	journal	November 2004
Sequence Determinants of Intracellular Phase Separation by Complex Coacervation of a Disordered Protein Pak, Chi W.; Kosno, Martyna; Holehouse, Alex S. Molecular Cell, Vol. 63, Issue 1 https://doi.org/10.1016/j.molcel.2016.05.042	journal	July 2016
How Hofmeister ion interactions affect protein stability Baldwin, R. L. Biophysical Journal, Vol. 71, Issue 4, p. 2056-2063 https://doi.org/10.1016/S0006-3495(96)79404-3	journal	October 1996
Atomic-Level Characterization of the Structural Dynamics of Proteins Shaw, D. E.; Maragakis, P.; Lindorff-Larsen, K. Science, Vol. 330, Issue 6002 https://doi.org/10.1126/science.1187409	journal	October 2010
Fast Parallel Algorithms for Short-Range Molecular Dynamics Plimpton, Steve Journal of Computational Physics, Vol. 117, Issue 1 https://doi.org/10.1006/jcph.1995.1039	journal	March 1995
Phosphorylation-Regulated Binding of RNA Polymerase II to Fibrous Polymers of Low-Complexity Domains Kwon, Ilmin; Kato, Masato; Xiang, Siheng Cell, Vol. 155, Issue 5 https://doi.org/10.1016/j.cell.2013.10.033	journal	November 2013
Generalized Replica Exchange Method Kim, Jaegil; Keyes, Thomas; Straub, John E. The Journal of Chemical Physics, Vol. 132, Issue 22 https://doi.org/10.1063/1.3432176	journal	June 2010
Conformational Heterogeneity and FRET Data Interpretation for Dimensions of Unfolded Proteins Song, Jianhui; Gomes, Gregory-Neal; Shi, Tongfei Biophysical Journal, Vol. 113, Issue 5 https://doi.org/10.1016/j.bpj.2017.07.023	journal	September 2017
Probing the Action of Chemical Denaturant on an Intrinsically Disordered Protein by Simulation and Experiment Zheng, Wenwei; Borgia, Alessandro; Buholzer, Karin Journal of the American Chemical Society, Vol. 138, Issue 36 https://doi.org/10.1021/jacs.6b05443	journal	September 2016
Phase Transitions in Biological Systems with Many Components Jacobs, William M.; Frenkel, Daan Biophysical Journal, Vol. 112, Issue 4 https://doi.org/10.1016/j.bpj.2016.10.043	journal	February 2017
Protein intrinsic disorder-based liquid–liquid phase transitions in biological systems: Complex coacervates and membrane-less organelles Uversky, Vladimir N. Advances in Colloid and Interface Science, Vol. 239 https://doi.org/10.1016/j.cis.2016.05.012	journal	January 2017
Polymer-Polymer Phase Behavior Bates, Frank S. Science, Vol. 251, Issue 4996, p. 898-905 https://doi.org/10.1126/science.251.4996.898	journal	February 1991
Random-phase-approximation theory for sequence-dependent, biologically functional liquid-liquid phase separation of intrinsically disordered proteins Lin, Yi-Hsuan; Song, Jianhui; Forman-Kay, Julie D. Journal of Molecular Liquids, Vol. 228 https://doi.org/10.1016/j.molliq.2016.09.090	journal	February 2017
Polymer scaling laws of unfolded and intrinsically disordered proteins quantified with single-molecule spectroscopy Hofmann, H.; Soranno, A.; Borgia, A. Proceedings of the National Academy of Sciences, Vol. 109, Issue 40 https://doi.org/10.1073/pnas.1207719109	journal	September 2012
Sequence heuristics to encode phase behaviour in intrinsically disordered protein polymers Quiroz, Felipe García; Chilkoti, Ashutosh Nature Materials, Vol. 14, Issue 11 https://doi.org/10.1038/nmat4418	journal	September 2015
Balanced Protein–Water Interactions Improve Properties of Disordered Proteins and Non-Specific Protein Association Best, Robert B.; Zheng, Wenwei; Mittal, Jeetain Journal of Chemical Theory and Computation, Vol. 10, Issue 11 https://doi.org/10.1021/ct500569b	journal	October 2014
Coarse-grained Models for Simulations of Multiprotein Complexes: Application to Ubiquitin Binding Kim, Young C.; Hummer, Gerhard Journal of Molecular Biology, Vol. 375, Issue 5 https://doi.org/10.1016/j.jmb.2007.11.063	journal	February 2008
A theoretical method to compute sequence dependent configurational properties in charged polymers and proteins Sawle, Lucas; Ghosh, Kingshuk The Journal of Chemical Physics, Vol. 143, Issue 8 https://doi.org/10.1063/1.4929391	journal	August 2015
Cell-free Formation of RNA Granules: Low Complexity Sequence Domains Form Dynamic Fibers within Hydrogels Kato, Masato; Han, Tina W.; Xie, Shanhai Cell, Vol. 149, Issue 4 https://doi.org/10.1016/j.cell.2012.04.017	journal	May 2012
Structural Basis for RNA Unwinding by the DEAD-Box Protein Drosophila Vasa Sengoku, Toru; Nureki, Osamu; Nakamura, Akira Cell, Vol. 125, Issue 2 https://doi.org/10.1016/j.cell.2006.01.054	journal	April 2006
General purpose molecular dynamics simulations fully implemented on graphics processing units Anderson, Joshua A.; Lorenz, Chris D.; Travesset, A. Journal of Computational Physics, Vol. 227, Issue 10 https://doi.org/10.1016/j.jcp.2008.01.047	journal	May 2008
Phase transitions in the assembly of multivalent signalling proteins Li, Pilong; Banjade, Sudeep; Cheng, Hui-Chun Nature, Vol. 483, Issue 7389 https://doi.org/10.1038/nature10879	journal	March 2012
Reversible Compartmentalization of de Novo Purine Biosynthetic Complexes in Living Cells An, S.; Kumar, R.; Sheets, E. D. Science, Vol. 320, Issue 5872 https://doi.org/10.1126/science.1152241	journal	April 2008
Residue – Residue Potentials with a Favorable Contact Pair Term and an Unfavorable High Packing Density Term, for Simulation and Threading Miyazawa, Sanzo; Jernigan, Robert L. Journal of Molecular Biology, Vol. 256, Issue 3 https://doi.org/10.1006/jmbi.1996.0114	journal	March 1996
A simple method for displaying the hydropathic character of a protein Kyte, Jack; Doolittle, Russell F. Journal of Molecular Biology, Vol. 157, Issue 1, p. 105-132 https://doi.org/10.1016/0022-2836(82)90515-0	journal	May 1982
Phase Transition of a Disordered Nuage Protein Generates Environmentally Responsive Membraneless Organelles Nott, Timothy J.; Petsalaki, Evangelia; Farber, Patrick Molecular Cell, Vol. 57, Issue 5 https://doi.org/10.1016/j.molcel.2015.01.013	journal	March 2015
Polymer physics of intracellular phase transitions Brangwynne, Clifford P.; Tompa, Peter; Pappu, Rohit V. Nature Physics, Vol. 11, Issue 11 https://doi.org/10.1038/nphys3532	journal	November 2015
Spatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets Shin, Yongdae; Berry, Joel; Pannucci, Nicole Cell, Vol. 168, Issue 1-2 https://doi.org/10.1016/j.cell.2016.11.054	journal	January 2017
Natively Unfolded Protein Stability as a Coil-to-Globule Transition in Charge/Hydropathy Space Ashbaugh, Henry S.; Hatch, Harold W. Journal of the American Chemical Society, Vol. 130, Issue 29 https://doi.org/10.1021/ja802124e	journal	July 2008
Coexisting Liquid Phases Underlie Nucleolar Subcompartments Feric, Marina; Vaidya, Nilesh; Harmon, Tyler S. Cell, Vol. 165, Issue 7 https://doi.org/10.1016/j.cell.2016.04.047	journal	June 2016
Protein Composition Determines the Effect of Crowding on the Properties of Disordered Proteins Miller, Cayla M.; Kim, Young C.; Mittal, Jeetain Biophysical Journal, Vol. 111, Issue 1 https://doi.org/10.1016/j.bpj.2016.05.033	journal	July 2016
Mutations in FUS, an RNA Processing Protein, Cause Familial Amyotrophic Lateral Sclerosis Type 6 Vance, C.; Rogelj, B.; Hortobagyi, T. Science, Vol. 323, Issue 5918 https://doi.org/10.1126/science.1165942	journal	February 2009
Nuclear Stress Bodies Biamonti, G.; Vourc'h, C. Cold Spring Harbor Perspectives in Biology, Vol. 2, Issue 6 https://doi.org/10.1101/cshperspect.a000695	journal	April 2010
Effects of salts on the free energies of nonpolar groups in model peptides Nandi, Pradip K.; Robinson, Dwight R. Journal of the American Chemical Society, Vol. 94, Issue 4 https://doi.org/10.1021/ja00759a043	journal	February 1972
Effects of salts on the free energy of the peptide group Nandi, Pradip K.; Robinson, Dwight R. Journal of the American Chemical Society, Vol. 94, Issue 4 https://doi.org/10.1021/ja00759a042	journal	February 1972
The Role of Nuclear Bodies in Gene Expression and Disease Morimoto, Marie; Boerkoel, Cornelius Biology, Vol. 2, Issue 3 https://doi.org/10.3390/biology2030976	journal	July 2013
Dual Specificity Kinase DYRK3 Couples Stress Granule Condensation/Dissolution to mTORC1 Signaling Wippich, Frank; Bodenmiller, Bernd; Trajkovska, Maria Gustafsson Cell, Vol. 152, Issue 4 https://doi.org/10.1016/j.cell.2013.01.033	journal	February 2013
A Liquid-to-Solid Phase Transition of the ALS Protein FUS Accelerated by Disease Mutation Patel, Avinash; Lee, Hyun O.; Jawerth, Louise Cell, Vol. 162, Issue 5 https://doi.org/10.1016/j.cell.2015.07.047	journal	August 2015
Membraneless organelles can melt nucleic acid duplexes and act as biomolecular filters Nott, Timothy J.; Craggs, Timothy D.; Baldwin, Andrew J. Nature Chemistry, Vol. 8, Issue 6 https://doi.org/10.1038/nchem.2519	journal	May 2016
Regulation of RNA granule dynamics by phosphorylation of serine-rich, intrinsically disordered proteins in C. elegans Wang, Jennifer T.; Smith, Jarrett; Chen, Bi-Chang eLife, Vol. 3 https://doi.org/10.7554/eLife.04591	journal	December 2014
Phosphorylation-Regulated Binding of RNA Polymerase II to Fibrous Polymers of Low-Complexity Domains Kwon, Ilmin; Kato, Masato; Xiang, Siheng Cell, Vol. 156, Issue 1-2 https://doi.org/10.1016/j.cell.2014.01.002	journal	January 2014
The Role of Nuclear Bodies in Gene Expression and Disease Morimoto, Marie; Boerkoel, Cornelius F. Multidisciplinary Digital Publishing Institute https://doi.org/10.14288/1.0378029	text	January 2019
Phase Transitions in Biological Systems with Many Components Jacobs, William M.; Frenkel, Daan Apollo - University of Cambridge Repository https://doi.org/10.17863/cam.17494	text	January 2017
Probing the Action of Chemical Denaturant on an Intrinsically Disordered Protein by Simulation and Experiment Zheng, Wenwei; Borgia, Alessandro; Buholzer, Karin American Chemical Society (ACS) https://doi.org/10.5167/uzh-126516	text	January 2016
Consistent view of polypeptide chain expansion in chemical denaturants from multiple experimental methods Borgia, Alessandro; Zheng, Wenwei; Buholzer, Karin American Chemical Society (ACS) https://doi.org/10.5167/uzh-126517	text	January 2016
Polymer scaling laws of unfolded and intrinsically disordered proteins quantified with single-molecule spectroscopy Hofmann, Hagen; Soranno, Andrea; Borgia, Alessandro National Academy of Sciences https://doi.org/10.5167/uzh-65974	text	January 2012
Dual specificity kinase DYRK3 couples stress granule condensation/dissolution to mTORC1 signaling Wippich, Frank; Bodenmiller, Bernd; Trajkovska, Maria Gustafsson Cell Press (Elsevier) https://doi.org/10.5167/uzh-77309	text	January 2013
Thermodynamics of High Polymer Solutions Flory, Paul J. The Journal of Chemical Physics, Vol. 9, Issue 8 https://doi.org/10.1063/1.1750971	journal	August 1941
How accurate are polymer models in the analysis of Forster resonance energy transfer experiments on proteins? O'Brien, E. P.; Morrison, G.; Brooks, B. R. arXiv https://doi.org/10.48550/arxiv.0905.0383	text	January 2009
Sequence-specific polyampholyte phase separation in membraneless organelles Lin, Yi-Hsuan; Forman-Kay, Julie D.; Chan, Hue Sun arXiv https://doi.org/10.48550/arxiv.1605.09019	text	January 2016
Random-phase-approximation theory for sequence-dependent, biologically functional liquid-liquid phase separation of intrinsically disordered proteins Lin, Yi-Hsuan; Song, Jianhui; Forman-Kay, Julie D. arXiv https://doi.org/10.48550/arxiv.1609.08205	text	January 2016
In Vitro Reconstitution of a Cellular Phase-Transition Process that Involves the mRNA Decapping Machinery Fromm, Simon A.; Kamenz, Julia; Nöldeke, Erik R. Angewandte Chemie International Edition, Vol. 53, Issue 28 https://doi.org/10.1002/anie.201402885	journal	May 2014
Comparative Protein Modelling by Satisfaction of Spatial Restraints Šali, Andrej; Blundell, Tom L. Journal of Molecular Biology, Vol. 234, Issue 3 https://doi.org/10.1006/jmbi.1993.1626	journal	December 1993
A simple method for displaying the hydropathic character of a protein Kyte, Jack; Doolittle, Russell F. Journal of Molecular Biology, Vol. 157, Issue 1, p. 105-132 https://doi.org/10.1016/0022-2836(82)90515-0	journal	May 1982
Protein Composition Determines the Effect of Crowding on the Properties of Disordered Proteins Miller, Cayla M.; Kim, Young C.; Mittal, Jeetain Biophysical Journal, Vol. 111, Issue 1 https://doi.org/10.1016/j.bpj.2016.05.033	journal	July 2016
To Mix, or To Demix, That Is the Question Harmon, Tyler S.; Holehouse, Alex S.; Pappu, Rohit V. Biophysical Journal, Vol. 112, Issue 4 https://doi.org/10.1016/j.bpj.2016.12.031	journal	February 2017
SAXS versus FRET: A Matter of Heterogeneity? Ruff, Kiersten M.; Holehouse, Alex S. Biophysical Journal, Vol. 113, Issue 5 https://doi.org/10.1016/j.bpj.2017.07.024	journal	September 2017
Structural Basis for RNA Unwinding by the DEAD-Box Protein Drosophila Vasa Sengoku, Toru; Nureki, Osamu; Nakamura, Akira Cell, Vol. 125, Issue 2 https://doi.org/10.1016/j.cell.2006.01.054	journal	April 2006
Bent out of Shape: RNA Unwinding by the DEAD-Box Helicase Vasa Linder, Patrick; Lasko, Paul Cell, Vol. 125, Issue 2 https://doi.org/10.1016/j.cell.2006.03.030	journal	April 2006
Cell-free Formation of RNA Granules: Low Complexity Sequence Domains Form Dynamic Fibers within Hydrogels Kato, Masato; Han, Tina W.; Xie, Shanhai Cell, Vol. 149, Issue 4 https://doi.org/10.1016/j.cell.2012.04.017	journal	May 2012
Dual Specificity Kinase DYRK3 Couples Stress Granule Condensation/Dissolution to mTORC1 Signaling Wippich, Frank; Bodenmiller, Bernd; Trajkovska, Maria Gustafsson Cell, Vol. 152, Issue 4 https://doi.org/10.1016/j.cell.2013.01.033	journal	February 2013
Phase Transition of Spindle-Associated Protein Regulate Spindle Apparatus Assembly Jiang, Hao; Wang, Shusheng; Huang, Yuejia Cell, Vol. 163, Issue 1 https://doi.org/10.1016/j.cell.2015.08.010	journal	September 2015
Phase Separation by Low Complexity Domains Promotes Stress Granule Assembly and Drives Pathological Fibrillization Molliex, Amandine; Temirov, Jamshid; Lee, Jihun Cell, Vol. 163, Issue 1 https://doi.org/10.1016/j.cell.2015.09.015	journal	September 2015
The LC Domain of hnRNPA2 Adopts Similar Conformations in Hydrogel Polymers, Liquid-like Droplets, and Nuclei Xiang, Siheng; Kato, Masato; Wu, Leeju C. Cell, Vol. 163, Issue 4 https://doi.org/10.1016/j.cell.2015.10.040	journal	November 2015
Coexisting Liquid Phases Underlie Nucleolar Subcompartments Feric, Marina; Vaidya, Nilesh; Harmon, Tyler S. Cell, Vol. 165, Issue 7 https://doi.org/10.1016/j.cell.2016.04.047	journal	June 2016
Spatiotemporal Control of Intracellular Phase Transitions Using Light-Activated optoDroplets Shin, Yongdae; Berry, Joel; Pannucci, Nicole Cell, Vol. 168, Issue 1-2 https://doi.org/10.1016/j.cell.2016.11.054	journal	January 2017
Gel or Die: Phase Separation as a Survival Strategy Kroschwald, Sonja; Alberti, Simon Cell, Vol. 168, Issue 6 https://doi.org/10.1016/j.cell.2017.02.029	journal	March 2017
Intrinsically disordered proteins as crucial constituents of cellular aqueous two phase systems and coacervates Uversky, Vladimir N.; Kuznetsova, Irina M.; Turoverov, Konstantin K. FEBS Letters, Vol. 589, Issue 1 https://doi.org/10.1016/j.febslet.2014.11.028	journal	November 2014
Coarse-grained Models for Simulations of Multiprotein Complexes: Application to Ubiquitin Binding Kim, Young C.; Hummer, Gerhard Journal of Molecular Biology, Vol. 375, Issue 5 https://doi.org/10.1016/j.jmb.2007.11.063	journal	February 2008
Residue-by-Residue View of In Vitro FUS Granules that Bind the C-Terminal Domain of RNA Polymerase II Burke, Kathleen A.; Janke, Abigail M.; Rhine, Christy L. Molecular Cell, Vol. 60, Issue 2 https://doi.org/10.1016/j.molcel.2015.09.006	journal	October 2015
Sequence Determinants of Intracellular Phase Separation by Complex Coacervation of a Disordered Protein Pak, Chi W.; Kosno, Martyna; Holehouse, Alex S. Molecular Cell, Vol. 63, Issue 1 https://doi.org/10.1016/j.molcel.2016.05.042	journal	July 2016
RNA Remodeling Activity of DEAD Box Proteins Tuned by Protein Concentration, RNA Length, and ATP Kim, Younghoon; Myong, Sua Molecular Cell, Vol. 63, Issue 5 https://doi.org/10.1016/j.molcel.2016.07.010	journal	September 2016
ALS Mutations Disrupt Phase Separation Mediated by α-Helical Structure in the TDP-43 Low-Complexity C-Terminal Domain Conicella, Alexander E.; Zerze, Gül H.; Mittal, Jeetain Structure, Vol. 24, Issue 9 https://doi.org/10.1016/j.str.2016.07.007	journal	September 2016
A New Phase in ALS Research Chong, P. Andrew; Forman-Kay, Julie D. Structure, Vol. 24, Issue 9 https://doi.org/10.1016/j.str.2016.08.003	journal	September 2016
The salting-out behavior of amides and its relation to the denaturation of proteins by salts Schrier, Eugene E.; Schrier, Evelyn B. The Journal of Physical Chemistry, Vol. 71, Issue 6 https://doi.org/10.1021/j100865a049	journal	May 1967
Effects of salts on the free energy of the peptide group Nandi, Pradip K.; Robinson, Dwight R. Journal of the American Chemical Society, Vol. 94, Issue 4 https://doi.org/10.1021/ja00759a042	journal	February 1972
Effects of salts on the free energies of nonpolar groups in model peptides Nandi, Pradip K.; Robinson, Dwight R. Journal of the American Chemical Society, Vol. 94, Issue 4 https://doi.org/10.1021/ja00759a043	journal	February 1972
Natively Unfolded Protein Stability as a Coil-to-Globule Transition in Charge/Hydropathy Space Ashbaugh, Henry S.; Hatch, Harold W. Journal of the American Chemical Society, Vol. 130, Issue 29 https://doi.org/10.1021/ja802124e	journal	July 2008
Probing the Action of Chemical Denaturant on an Intrinsically Disordered Protein by Simulation and Experiment Zheng, Wenwei; Borgia, Alessandro; Buholzer, Karin Journal of the American Chemical Society, Vol. 138, Issue 36 https://doi.org/10.1021/jacs.6b05443	journal	September 2016
Consistent View of Polypeptide Chain Expansion in Chemical Denaturants from Multiple Experimental Methods Borgia, Alessandro; Zheng, Wenwei; Buholzer, Karin Journal of the American Chemical Society, Vol. 138, Issue 36 https://doi.org/10.1021/jacs.6b05917	journal	September 2016
Water Dispersion Interactions Strongly Influence Simulated Structural Properties of Disordered Protein States Piana, Stefano; Donchev, Alexander G.; Robustelli, Paul The Journal of Physical Chemistry B, Vol. 119, Issue 16 https://doi.org/10.1021/jp508971m	journal	April 2015
Phase transitions in the assembly of multivalent signalling proteins Li, Pilong; Banjade, Sudeep; Cheng, Hui-Chun Nature, Vol. 483, Issue 7389 https://doi.org/10.1038/nature10879	journal	March 2012
Membraneless organelles can melt nucleic acid duplexes and act as biomolecular filters Nott, Timothy J.; Craggs, Timothy D.; Baldwin, Andrew J. Nature Chemistry, Vol. 8, Issue 6 https://doi.org/10.1038/nchem.2519	journal	May 2016
Phase behaviour of disordered proteins underlying low density and high permeability of liquid organelles Wei, Ming-Tzo; Elbaum-Garfinkle, Shana; Holehouse, Alex S. Nature Chemistry, Vol. 9, Issue 11 https://doi.org/10.1038/nchem.2803	journal	June 2017
Thermodynamics of High Polymer Solutions Flory, Paul J. The Journal of Chemical Physics, Vol. 10, Issue 1 https://doi.org/10.1063/1.1723621	journal	January 1942
Net charge per residue modulates conformational ensembles of intrinsically disordered proteins Mao, A. H.; Crick, S. L.; Vitalis, A. Proceedings of the National Academy of Sciences, Vol. 107, Issue 18 https://doi.org/10.1073/pnas.0911107107	journal	April 2010
Active liquid-like behavior of nucleoli determines their size and shape in Xenopus laevis oocytes Brangwynne, C. P.; Mitchison, T. J.; Hyman, A. A. Proceedings of the National Academy of Sciences, Vol. 108, Issue 11 https://doi.org/10.1073/pnas.1017150108	journal	February 2011
Polymer scaling laws of unfolded and intrinsically disordered proteins quantified with single-molecule spectroscopy Hofmann, H.; Soranno, A.; Borgia, A. Proceedings of the National Academy of Sciences, Vol. 109, Issue 40 https://doi.org/10.1073/pnas.1207719109	journal	September 2012
The disordered P granule protein LAF-1 drives phase separation into droplets with tunable viscosity and dynamics Elbaum-Garfinkle, Shana; Kim, Younghoon; Szczepaniak, Krzysztof Proceedings of the National Academy of Sciences, Vol. 112, Issue 23 https://doi.org/10.1073/pnas.1504822112	journal	May 2015
RNA transcription modulates phase transition-driven nuclear body assembly Berry, Joel; Weber, Stephanie C.; Vaidya, Nilesh Proceedings of the National Academy of Sciences, Vol. 112, Issue 38 https://doi.org/10.1073/pnas.1509317112	journal	September 2015
Decoupling of size and shape fluctuations in heteropolymeric sequences reconciles discrepancies in SAXS vs. FRET measurements Fuertes, Gustavo; Banterle, Niccolò; Ruff, Kiersten M. Proceedings of the National Academy of Sciences, Vol. 114, Issue 31 https://doi.org/10.1073/pnas.1704692114	journal	July 2017
Structural and hydrodynamic properties of an intrinsically disordered region of a germ cell-specific protein on phase separation Brady, Jacob P.; Farber, Patrick J.; Sekhar, Ashok Proceedings of the National Academy of Sciences, Vol. 114, Issue 39 https://doi.org/10.1073/pnas.1706197114	journal	September 2017
Nuclear Stress Bodies Biamonti, G.; Vourc'h, C. Cold Spring Harbor Perspectives in Biology, Vol. 2, Issue 6 https://doi.org/10.1101/cshperspect.a000695	journal	April 2010
Reversible Compartmentalization of de Novo Purine Biosynthetic Complexes in Living Cells An, S.; Kumar, R.; Sheets, E. D. Science, Vol. 320, Issue 5872 https://doi.org/10.1126/science.1152241	journal	April 2008
Mutations in the FUS/TLS Gene on Chromosome 16 Cause Familial Amyotrophic Lateral Sclerosis Kwiatkowski, T. J.; Bosco, D. A.; LeClerc, A. L. Science, Vol. 323, Issue 5918 https://doi.org/10.1126/science.1166066	journal	February 2009
Germline P Granules Are Liquid Droplets That Localize by Controlled Dissolution/Condensation Brangwynne, C. P.; Eckmann, C. R.; Courson, D. S. Science, Vol. 324, Issue 5935 https://doi.org/10.1126/science.1172046	journal	May 2009
Atomic-Level Characterization of the Structural Dynamics of Proteins Shaw, D. E.; Maragakis, P.; Lindorff-Larsen, K. Science, Vol. 330, Issue 6002 https://doi.org/10.1126/science.1187409	journal	October 2010
Polymer-Polymer Phase Behavior Bates, Frank S. Science, Vol. 251, Issue 4996, p. 898-905 https://doi.org/10.1126/science.251.4996.898	journal	February 1991
Innovative scattering analysis shows that hydrophobic disordered proteins are expanded in water Riback, Joshua A.; Bowman, Micayla A.; Zmyslowski, Adam M. Science, Vol. 358, Issue 6360 https://doi.org/10.1126/science.aan5774	journal	October 2017
An aberrant phase transition of stress granules triggered by misfolded protein and prevented by chaperone function Mateju, Daniel; Franzmann, Titus M.; Patel, Avinash The EMBO Journal, Vol. 36, Issue 12 https://doi.org/10.15252/embj.201695957	journal	April 2017
The Role of Nuclear Bodies in Gene Expression and Disease Morimoto, Marie; Boerkoel, Cornelius Biology, Vol. 2, Issue 3 https://doi.org/10.3390/biology2030976	journal	July 2013

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