DOCSLIB.ORG

Phase Separation in Biology

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Phase Separation in Biology Current Biology Magazine has not yet been applied to all Primer cytoplasm and the distribution of breeding programs, and there are these macromolecules is subject to still some major crop species (e.g., constant change. Early concepts to Phase separation soybean, tomato, sunfl ower) that are explain these density fl uctuations recalcitrant to the currently available in biology proposed a role for local synthesis, procedures, or for which present anchoring or active transport. Yet these protocols are not effi cient enough. Simon Alberti concepts are insuffi cient to explain The understanding of the maize in the complex organization of cells. situ system should help to extend Cells have to organize their complex We now realize that cells can only be the DH technology to more species biochemistry to regulate their properly understood if we also consider or breeding programs by either metabolism and respond to changes the collective properties of biological directly knocking out the functional in the environment. Traditionally, macromolecules. ortholog of the maize gene, or if intracellular organization has been Chemists have known for a long needed by transferring the cellular associated with compartments that time that macromolecules can have and molecular knowledge acquired are surrounded by lipid membranes. collective properties. The science of on maize. In addition to these However, in recent years, phase macromolecular collectives is called applications, the identifi cation of transitions have emerged as a novel colloidal chemistry. One example of a the NLD/MTL/ZmPLA1 gene offers form of cellular organization. Phase colloid is Jell-O, which is composed a unique opportunity to explore the transition is a physical process of colored water and gelatin, a large, many mysteries of double fertilization whereby a substance changes string-like protein. Gelatin dissolves in plants. from one physical state to another. well in hot water; however, when the Examples are provided by the freezing water is cooled, the gelatin molecules Where can I fi nd out more? of water into ice (liquid to solid) or the become insoluble and sticky and then Britt, A.B., and Kuppu, S. (2016). Cenh3: heating of water to generate water form a cross-linked network. The result an emerging player in haploid induction vapor (liquid to gas). of this process is a hydrogel. Technology. Front. Plant Sci. 7, 357. Dunwell, J.M. (2010). Haploids in fl owering plants: Liquid–liquid phase separation is The concept of viewing cells as origins and exploitation. Plant Biotechnol. J. a special form of a phase transition colloidal systems was fi rst proposed 8, 377–424. Dwivedi, S.L., Britt, A.B., Tripathi, L., Sharma, that is particularly relevant for living in the late 19th century by Edmund S., Upadhyaya, H.D., and Ortiz, R. (2015). organisms. Here, a homogenous Wilson. Wilson pointed out that cells Haploids: Constraints and opportunities in solution of molecules spontaneously appear as if they are densely packed plant breeding. Biotechnol. Adv. 33, 812–829. Forster, B.P., Heberle-Bors, E., Kasha, K.J., separates (‘demixes’) into two with macromolecular phases or liquid and Touraev, A. (2007). The resurgence of coexisting liquid phases, a dense phase ‘coacervates’. Alexander Oparin haploids in higher plants. Trends Plant Sci. 12, that is enriched for these molecules and later considered these coacervates 368–375. Gilles, L.M., Khaled, A., Laffaire, J.-B., a phase that is depleted (Figure 1). The an essential constituent of the Chaignon, S., Gendrot, G., Laplaige, J., interface of these dense droplets forms primordial soup. However, these initial Bergès, H., Beydon, G., Bayle, V., Barret, P., et al. (2017). Loss of pollen-specifi c a boundary that allows the selective concepts of cellular organization were phospholipase NOT LIKE DAD triggers passage of some molecules but not eclipsed by the enormous success of gynogenesis in maize. EMBO J. 36, 707–717. others. The presence of this interface reductionist molecular biology. Very Ishii, T., Karimi-Ashtiyani, R., and Houben, A. (2016). Haploidization via chromosome makes it possible that liquid droplets recently, however, we have seen a elimination: Means and mechanisms. Annu. can function as compartments. Once revival of colloidal biochemistry. Key Rev. Plant Biol. 67, 421–438. formed, liquid droplet compartments was the realization that cells contain Jackson, D. (2017). No sex please, we’re (in) breeding. EMBO J. 36, 703–704. can also adopt different physical states; compartments that lack membranes. Kelliher, T., Starr, D., Richbourg, L., Chintamanani, they can, for example, harden into gel- S., Delzer, B., Nuccio, M.L., Green, J., Chen, Z., McCuiston, J., Wang, W., et al. or glass-like states or they can turn into Membrane-less organelles (2017). MATRILINEAL, a sperm-specifi c solid crystals. Thus, phase separation Membrane-less organelles are micron- phospholipase, triggers maize haploid allows the formation of a wide range of sized cellular structures that consist of induction. Nature 542, 105–109. Liu, C., Li, X., Meng, D., Zhong, Y., Chen, C., compartments with different physical large collections of macromolecules, Dong, X., Xu, X., Chen, B., Li, W., Li, L., et al. properties. such as RNAs and proteins. Modern (2017). A 4-bp insertion at ZmPLA1 encoding a putative phospholipase A generates haploid In this Primer, I introduce the imaging approaches are revealing an induction in maize. Mol. Plant 10, 520–522. emerging topic of biological phase increasing number of these structures Ren, J., Wu, P., Trampe, B., Tian, X., transitions. I further discuss how cells (Figure 2). One prominent example is Lübberstedt, T., and Chen, S. (2017). Novel technologies in doubled haploid line use phase transitions to regulate provided by the centrosome, which development. Plant Biotechnol. J., http:// biological functions and activities in organizes the microtubule network of dx.doi.org/10.1111/pbi.1280. order to increase their fi tness and cells. Another example is provided by chances of survival. processing bodies or P bodies, which 1Laboratoire Reproduction et store and degrade RNA. Then there Dé veloppement des Plantes, Univ Lyon, The colloidal nature of cells is the example of the nucleolus, an ENS de Lyon, UCB Lyon 1, CNRS, INRA, The cellular interior is not a well- assembly line for ribosomes: it has a F-69342, Lyon, France; 2Limagrain, Limagrain Field Seeds, Research Centre, mixed collection of molecules. fi brillar center, where the ribosomal F-63360 Gerzat, France. Macromolecules are enriched or RNA is produced, and two additional *E-mail: [email protected] depleted in certain areas of the intranucleolar compartments, which Current Biology 27, R1089–R1107, October 23, 2017 © 2017 Elsevier Ltd. R1097 Current Biology Magazine In a key paper in 2012, Mike Phase separation Protein Rosen and colleagues provided conc. the fi rst experimental evidence that multivalency — the availability of many different binding sites on a molecule — is the crucial parameter that determines whether or not a protein will undergo phase separation. C Critical Many proteins, especially those functioning in signaling or gene expression, contain tandem arrays of modular domains. Each of these interaction modules binds a particular Time ligand, often with relatively weak affi nity. The number of these different modules determines the valency of Protein Phase separation the molecule. Importantly, multivalent conc. proteins have a critical threshold for phase separation that depends on the number of modules and available ligands. Increasing the number of the modules promotes the formation of increasingly larger complexes that CCritical PTMs, temperature, manifest macroscopically as phase- ionic strength separated droplets. Thus, although the C Critical composition of cellular bodies appears to be complex, their formation may be governed by very simple rules, such as Time protein valency and the availability of Current Biology ligands. Another feature that strongly affects Figure 1. Protein phase separation. the phase behavior of proteins is their Top: A protein forms dense liquid droplets above the critical concentration (Ccritical) for phase separa- solubility. Many phase-separating tion. The liquid droplets are stable as long as the total concentration is above the critical threshold. proteins have poor solubility in water. The droplets form a compartment that allows the diffusion of molecules within the compartment and Burying such insoluble proteins in a promotes the dynamic exchange of molecules with the dilute surrounding phase. Once the total protein concentration drops below the critical concentration again, the compartments dissolve and the system growing phase-separating structure relapses back to a one-phase state where the molecules are evenly distributed. The insets above show is energetically more favorable than original data from a phase separation experiment with purifi ed GFP-tagged FUS (a prion-like RNA- exposing these proteins to water. As binding protein). Bottom: Post-translational modifi cations (PTMs) or changes in temperature or ionic a consequence, solubility changes strength can increase the affi nity of protein–protein interactions, thus lowering the critical threshold for and phase separation are often phase separation and allowing compartment formation at a much lower protein concentration. coupled. This is particularly relevant for multidomain proteins that are assemble ribosomes from molecular protein–nucleic acid interactions connected
Load more

Recommended publications
  • Liquid Network Connectivity Regulates the Stability and Composition Of
    Liquid network connectivity regulates the stability and composition of biomolecular condensates with many components Jorge R. Espinosaa,b,c, Jerelle A. Josepha,b,c , Ignacio Sanchez-Burgosa,b,c , Adiran Garaizara,b,c , Daan Frenkelb , and Rosana Collepardo-Guevaraa,b,c,1 aMaxwell Centre, Cavendish Laboratory, Department of Physics, University of Cambridge, Cambridge CB3 0HE, United Kindgdom; bDepartment of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom; and cDepartment of Genetics, University of Cambridge, Cambridge CB2 3EH, United Kingdom Edited by Ken A. Dill, Stony Brook University, Stony Brook, NY, and approved April 17, 2020 (received for review October 8, 2019) One of the key mechanisms used by cells to control the spatiotem- 10), cellular bodies (11), or otherwise (8)—are ubiquitous within poral organization of their many components is the formation and the cell interior. Biomolecular condensates account for numer- dissolution of biomolecular condensates through liquid–liquid ous highly diverse domains, both inside the cytoplasm [e.g., P phase separation (LLPS). Using a minimal coarse-grained model granules (12) and RNA granules/bodies (13–15)] and in the cell that allows us to simulate thousands of interacting multivalent nucleus [e.g., Cajal bodies (16), nucleoli (17), heterochromatin proteins, we investigate the physical parameters dictating the domains (18, 19), the transport channels in the nuclear pore stability and composition of multicomponent biomolecular con- complex (20), and possibly superenhancers
    [Show full text]
  • S41598-020-74080-2.Pdf
    www.nature.com/scientificreports OPEN Multivalent tumor suppressor adenomatous polyposis coli promotes Axin biomolecular condensate formation and efcient β‑catenin degradation Tie‑Mei Li1,2,5*, Jing Ren3, Dylan Husmann2, John P. Coan1,2, Or Gozani2* & Katrin F. Chua1,4* The tumor suppressor adenomatous polyposis coli (APC) is frequently mutated in colorectal cancers. APC and Axin are core components of a destruction complex that scafolds GSK3β and CK1 to earmark β‑catenin for proteosomal degradation. Disruption of APC results in pathologic stabilization of β‑catenin and oncogenesis. However, the molecular mechanism by which APC promotes β‑catenin degradation is unclear. Here, we fnd that the intrinsically disordered region (IDR) of APC, which contains multiple β‑catenin and Axin interacting sites, undergoes liquid–liquid phase separation (LLPS) in vitro. Expression of the APC IDR in colorectal cells promotes Axin puncta formation and β‑catenin degradation. Our results support the model that multivalent interactions between APC and Axin drives the β‑catenin destruction complex to form biomolecular condensates in cells, which concentrate key components to achieve high efcient degradation of β‑catenin. APC mutations are present in ~ 80% of colorectal cancer cases1, and typically cause truncation of the APC pro- tein. APC functions downstream of the Wnt signalosome, and it is essential for the degradation of β-catenin in the absence of Wnt stimulation 2. APC forms a complex, termed the “β-catenin destruction complex” or “Axin degradasome” composed of β-catenin, the scafold protein Axin, and two kinases: GSK3β and casein kinase 1 (CK1)3,4. In the complex, proximity of β-catenin to the two kinases leads to β-catenin phosphorylation, which in turn facilitates its ubiquitination and proteosomal degradation.
    [Show full text]
  • Structure of Biomolecular Condensates from Dissipative Particle Dynamics Simulations
    bioRxiv preprint doi: https://doi.org/10.1101/2019.12.11.873133; this version posted April 30, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Structure of biomolecular condensates from dissipative particle dynamics simulations Julian C. Shillcock,1* Maelick Brochut,2 Etienne Chénais,2 and John H. Ipsen3 1 Laboratory of Molecular and Chemical Biology of Neurodegeneration, Ecole polytechnique fédérale de Lausanne, CH-1015 Lausanne, Switzerland 2 Brain Mind Institute, Ecole polytechnique fédérale de Lausanne, CH-1015 Lausanne, Switzerland 3 Dept. of Physics, Chemistry and Pharmacy, University of Southern Denmark, Campusvej 55, DK-5230 Odense M, Denmark * Correspondence: [email protected], OrcId: 0000-0002-7885-735X ABSTRACT Phase separation of immiscible fluids is a common phenomenon in polymer chemistry, and is recognized as an important mechanism by which cells compartmentalize their biochemical reactions. Biomolecular condensates are condensed fluid droplets in cells that form by liquid-liquid phase separation of intrinsically-disordered proteins. They have a wide range of functions and are associated with chronic neurodegenerative diseases in which they become pathologically rigid. Intrinsically-disordered proteins are conformationally flexible and possess multiple, distributed binding sites for each other or for RNA. However, it remains unclear how their material properties depend on the molecular structure of the proteins. Here we use coarse-grained simulations to explore the phase behavior and structure of a model biomolecular condensate composed of semi-flexible polymers with attractive end-caps in a good solvent. Although highly simplified, the model contains the minimal molecular features that are sufficient to observe liquid-liquid phase separation of soluble polymers.
    [Show full text]
  • The Nucleolus As a Multiphase Liquid Condensate
    REVIEWS The nucleolus as a multiphase liquid condensate Denis L. J. Lafontaine 1 ✉ , Joshua A. Riback 2, Rümeyza Bascetin 1 and Clifford P. Brangwynne 2,3 ✉ Abstract | The nucleolus is the most prominent nuclear body and serves a fundamentally important biological role as a site of ribonucleoprotein particle assembly, primarily dedicated to ribosome biogenesis. Despite being one of the first intracellular structures visualized historically, the biophysical rules governing its assembly and function are only starting to become clear. Recent studies have provided increasing support for the concept that the nucleolus represents a multilayered biomolecular condensate, whose formation by liquid–liquid phase separation (LLPS) facilitates the initial steps of ribosome biogenesis and other functions. Here, we review these biophysical insights in the context of the molecular and cell biology of the nucleolus. We discuss how nucleolar function is linked to its organization as a multiphase condensate and how dysregulation of this organization could provide insights into still poorly understood aspects of nucleolus-associated diseases, including cancer, ribosomopathies and neurodegeneration as well as ageing. We suggest that the LLPS model provides the starting point for a unifying quantitative framework for the assembly, structural maintenance and function of the nucleolus, with implications for gene regulation and ribonucleoprotein particle assembly throughout the nucleus. The LLPS concept is also likely useful in designing new therapeutic strategies to target nucleolar dysfunction. Protein trans-acting factors Among numerous microscopically visible nuclear sub- at the inner core where rRNA transcription occurs and Proteins important for structures, the nucleolus is the most prominent and proceeding towards the periphery (Fig.
    [Show full text]
  • Clusters of Bacterial RNA Polymerase Are Biomolecular Condensates That Assemble Through Liquid–Liquid Phase Separation
    Clusters of bacterial RNA polymerase are biomolecular condensates that assemble through liquid–liquid phase separation Anne-Marie Ladouceura,1, Baljyot Singh Parmarb,1, Stefan Biedzinskia, James Walla, S. Graydon Topea, David Cohna, Albright Kima, Nicolas Soubrya, Rodrigo Reyes-Lamothea, and Stephanie C. Webera,b,2 aDepartment of Biology, McGill University, Montreal, QC H3A 1B1, Canada; and bDepartment of Physics, McGill University, Montreal, QC H3A 2T8, Canada Edited by Richard A. Young, Massachusetts Institute of Technology, Cambridge, MA, and approved June 23, 2020 (received for review March 17, 2020) Once described as mere “bags of enzymes,” bacterial cells are in possibility that prokaryotes also use LLPS to compartmentalize fact highly organized, with many macromolecules exhibiting non- their cells. uniform localization patterns. Yet the physical and biochemical In eukaryotes, LLPS is mediated by proteins containing mul- mechanisms that govern this spatial heterogeneity remain largely tivalent domains and disordered regions (22–24) that bring unknown. Here, we identify liquid–liquid phase separation (LLPS) as molecules together into dynamic condensates through transient a mechanism for organizing clusters of RNA polymerase (RNAP) in interactions. Sequence analysis predicts that bacterial proteomes Escherichia coli . Using fluorescence imaging, we show that RNAP have a much lower frequency of disordered regions than quickly transitions from a dispersed to clustered localization pattern eukaryotic proteomes: Only 4.2% compared to 33%, respectively as cells enter log phase in nutrient-rich media. RNAP clusters are (25). This paucity of disorder raises doubts about the prevalence sensitive to hexanediol, a chemical that dissolves liquid-like com- of LLPS in this domain of life.
    [Show full text]
  • 2021.02.03.429226V1.Full.Pdf
    bioRxiv preprint doi: https://doi.org/10.1101/2021.02.03.429226; this version posted February 3, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. A modular platform for engineering function of natural and synthetic biomolecular condensates Keren Lasker1,*, Steven Boeynaems2,*, Vinson Lam3, Emma Stainton1, Maarten Jacquemyn4, Dirk Daelemans4, Elizabeth Villa3, Alex S. Holehouse5,6, Aaron D. Gitler2,#, Lucy Shapiro1,# 1Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA 94305, USA 2Department of Genetics, Stanford University School of Medicine, Stanford, California 94305, USA 3Division of Biological Sciences, University of California San Diego, La Jolla, CA 92093, USA 4KU Leuven Department of Microbiology, Immunology, and Transplantation, Laboratory of Virology and Chemotherapy, Rega Institute, KU Leuven, 3000 Leuven, Belgium 5Department of Biochemistry and Molecular Biophysics, Washington University in St. Louis, St. Louis, MO 63110, USA. 6Center for Science and Engineering of Living Systems (CSELS), Washington University in St. Louis, St. Louis, MO 63130, USA *Equal contribution, #emails: [email protected], [email protected] Abstract Phase separation is emerging as a universal principle for how cells use dynamic subcompartmentalization to organize biochemical reactions in time and space1,2. Yet, whether the emergent physical properties of these biomolecular condensates are important
    [Show full text]
  • Phase Separation in Biology and Disease—
    Ann. N.Y. Acad. Sci. ISSN 0077-8923 ANNALS OF THE NEW YORK ACADEMY OF SCIENCES Special Issue: Annals Reports COMMENTARY Phase separation in biology and disease—a symposium report Jennifer Cable,1 Clifford Brangwynne,2 Geraldine Seydoux,3 David Cowburn,4 Rohit V. Pappu,5 Carlos A. Castañeda,6 Luke E. Berchowitz,7 Zhijuan Chen,8 Martin Jonikas,9 Abby Dernburg,10 Tanja Mittag,11 and Nicolas L. Fawzi12 1Science Writer, New York, New York. 2Department of Chemical and Biological Engineering, Princeton University, Princeton, New Jersey. 3Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland. 4Departments of Biochemistry and Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, New York. 5Department of Biomedical Engineering, Center for Science and Engineering of Living Systems, McKelvey School of Engineering, St. Louis, Missouri. 6Departments of Biology and Chemistry, Program in Neuroscience, Syracuse University, Syracuse, New York. 7Departments of Genetics and Development, Columbia University, New York, New York. 8University of Texas Southwestern Medical Center, Dallas, Texas. 9Department of Molecular Biology, Princeton University, Princeton, New Jersey. 10Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California. 11Department of Structural Biology, St. Jude Children’s Research Hospital, Memphis, Tennessee. 12Department of Molecular Pharmacology, Physiology and Biotechnology, Brown University, Providence, Rhode Island Address for correspondence: [email protected] Phase separation of multivalent protein and RNA molecules enables cells the formation of reversible nonstoichio- metric, membraneless assemblies. These assemblies, referred to as biomolecular condensates, help with the spatial organization and compartmentalization of cellular matter. Each biomolecular condensate is defined by a distinct macromolecular composition.
    [Show full text]
  • New Perspectives on the Biogenesis of Viral Inclusion Bodies in Negative-Sense RNA Virus Infections
    cells Review New Perspectives on the Biogenesis of Viral Inclusion Bodies in Negative-Sense RNA Virus Infections Olga Dolnik , Gesche K. Gerresheim and Nadine Biedenkopf * Institute for Virology, Philipps-University Marburg, 35043 Marburg, Germany; [email protected] (O.D.); [email protected] (G.K.G.) * Correspondence: [email protected]; +49-(0)-64212825307 Abstract: Infections by negative strand RNA viruses (NSVs) induce the formation of viral inclusion bodies (IBs) in the host cell that segregate viral as well as cellular proteins to enable efficient viral replication. The induction of those membrane-less viral compartments leads inevitably to structural remodeling of the cellular architecture. Recent studies suggested that viral IBs have properties of biomolecular condensates (or liquid organelles), as have previously been shown for other membrane- less cellular compartments like stress granules or P-bodies. Biomolecular condensates are highly dynamic structures formed by liquid-liquid phase separation (LLPS). Key drivers for LLPS in cells are multivalent protein:protein and protein:RNA interactions leading to specialized areas in the cell that recruit molecules with similar properties, while other non-similar molecules are excluded. These typical features of cellular biomolecular condensates are also a common characteristic in the biogenesis of viral inclusion bodies. Viral IBs are predominantly induced by the expression of the viral nucleoprotein (N, NP) and phosphoprotein (P); both are characterized by a special protein architecture containing multiple disordered regions and RNA-binding domains that contribute to Citation: Dolnik, O.; Gerresheim, different protein functions. P keeps N soluble after expression to allow a concerted binding of N to G.K.; Biedenkopf, N.
    [Show full text]
  • RNA Impacts Formation of Biomolecular Condensates in the Nucleus
    Biomedical Research (Tokyo) 42 (4) 153–160, 2021 RNA impacts formation of biomolecular condensates in the nucleus Saho MATSUI and Ryu-Suke NOZAWA Division of Experimental Pathology, Cancer Institute of the Japanese Foundation for Cancer Research (JFCR), Ariake 3-8-31 Koto-ku, 135-8550 Tokyo, Japan (Received 10 May 2021; and accepted 11 June 2021) ABSTRACT Biomolecular condensates are membrane-less compartments that are formed through an assembly of proteins and nucleic acids in the cell. Dysregulation of biological condensates has been impli- cated in diseases such as neurodegeneration and cancer. Ribonucleic acid (RNA) is known to af- fect the assembly of proteins in vitro, if and how RNA is involved in regulating biomolecular condensates in cells is not well investigated. Here we examined two nuclear proteins, FUS and HP1α, in which RNA was found to have an opposite contribution for the assembly of these pro- teins. Reduction of nuclear RNA, by inhibiting the transcription, triggered assembly of FUS that had been distributed in the nucleoplasm, whereas it dispersed spontaneously formed HP1α assem- bly. Notably, the cell cycle-dependent phosphorylation-mimicking substitutions in HP1α promoted its assembly formation. These transcription inhibitor experiments are versatile to examine diverse roles of nuclear RNA in regulating biomolecular condensates, in both physiological and pathologi- cal conditions. uid phase composed of multiple soluble ingredients INTRODUCTION separating into different phases through their selec- Cells organize compartments in their interior to dif- tive and exclusive interactions (as a salad dressing ferentiate and facilitate biological processes. The nu- separates into oil- and water-phases). Through an cleus and mitochondria contain a lipid bilayer to analogous mechanism, selective assembly of bio- physically separate inside and outside and maintain molecules, such as proteins and nucleic acids, drives their internal biochemical environment.
    [Show full text]
  • Exploring Relationships Between the Density of Charged Tracts Within Disordered Regions and Phase Separation
    Pacific Symposium on Biocomputing 25:207-218(2020) Exploring Relationships between the Density of Charged Tracts within Disordered Regions and Phase Separation Ramiz Somjee Department of Structural Biology, St. Jude Children’s Research Hospital, 262 Danny Thomas Place Memphis, Tennessee 38105, USA; Department of Chemistry, Rhodes College, 2000 North Parkway Memphis, Tennessee 38112, USA; Email: [email protected] Diana M. Mitrea Department of Structural Biology, St. Jude Children’s Research Hospital, 262 Danny Thomas Place Memphis, Tennessee 38105, USA Email: [email protected] Richard W. Kriwacki† Department of Structural Biology, St. Jude Children’s Research Hospital, 262 Danny Thomas Place Memphis, Tennessee 38105, USA; Department of Microbiology, Immunology, and Biochemistry, University of Tennessee Health Sciences Center, 910 Madison Avenue, Memphis, Tennessee 38163, USA; Email: [email protected] Biomolecular condensates form through a process termed phase separation and play diverse roles throughout the cell. Proteins that undergo phase separation often have disordered regions that can engage in weak, multivalent interactions; however, our understanding of the sequence grammar that defines which proteins phase separate is far from complete. Here, we show that proteins that display a high density of charged tracts within intrinsically disordered regions are likely to be constituents of electrostatically organized biomolecular condensates. We scored the human proteome using an algorithm termed ABTdensity that quantifies the density of charged tracts and observed that proteins with more charged tracts are enriched in particular Gene Ontology annotations and, based upon analysis of interaction networks, cluster into distinct biomolecular condensates. These results suggest that electrostatically-driven, multivalent interactions involving charged tracts within disordered regions serve to organize certain biomolecular condensates through phase separation.
    [Show full text]
  • Programmable Viscoelasticity in Protein-RNA Condensates with Disordered Sticker- Spacer Polypeptides
    bioRxiv preprint doi: https://doi.org/10.1101/2021.01.24.427968; this version posted August 11, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Programmable Viscoelasticity in Protein-RNA Condensates with Disordered Sticker- Spacer Polypeptides Ibraheem Alshareedah1, Mahdi Muhammad Moosa1, Matthew Pham2, Davit A. Potoyan2*, Priya R. Banerjee1* 1Department of Physics, University at Buffalo, Buffalo NY 14260, USA 2Department of Chemistry, Iowa State University, Ames IA 50011, USA *All correspondence could be addressed to: Priya R. Banerjee (email: [email protected]) Davit A. Potoyan (email: [email protected]) bioRxiv preprint doi: https://doi.org/10.1101/2021.01.24.427968; this version posted August 11, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. ABSTRACT Liquid-liquid phase separation of multivalent proteins and RNAs drives the formation of biomolecular condensates that facilitate membrane-free compartmentalization of subcellular processes. With recent advances, it is becoming increasingly clear that biomolecular condensates are network fluids with time-dependent material properties. Here, employing microrheology with optical tweezers, we reveal molecular determinants that govern the viscoelastic behavior of condensates formed by multivalent Arg/Gly-rich sticker-spacer polypeptides and RNA. These condensates behave as Maxwell fluids with an elastically-dominant rheological response at shorter timescales and a liquid-like behavior at longer timescales.
    [Show full text]
  • 1 Emergent Functions of Proteins in Non-Stoichiometric Supramolecular
    *REVISEDView metadata, Manuscript citation and (textsimilar UNmarked) papers at core.ac.uk brought to you by CORE Click here to view linked References provided by Repository of the Academy's Library 1 Emergent functions of proteins in non-stoichiometric supramolecular assemblies Rita Pancsa1, Eva Schad1, Agnes Tantos1, Peter Tompa1,2,3 1Institute of Enzymology, Research Centre for Natural Sciences of the Hungarian Academy of Sciences, Budapest, Hungary 2VIB Center for Structural Biology (CSB), Brussels, Belgium 3Structural Biology Brussels (SBB), Vrije Universiteit Brussel (VUB), Brussels, Belgium, 2 Abstract Proteins are the basic functional units of the cell, carrying out myriads of functions essential for life. There are countless reports in molecular cell biology addressing the functioning of proteins under physiological and pathological conditions, aiming to understand life at the atomistic-molecular level and thereby being able to develop remedies against diseases. The central theme in most of these studies is that the functional unit under study is the protein itself. Recent rapid progress has radically challenged and extended this protein-function paradigm, by demonstrating that novel function(s) may emerge when proteins form dynamic and non-stoichiometric supramolecular assemblies. There is an increasing number of cases for such collective functions, such as targeting, localization, protection/shielding and filtering effects, as exemplified by signaling complexes and prions, biominerals and mucus, amphibian adhesions and bacterial biofilms, and a broad range of membraneless organelles (bio-condensates) formed by liquid-liquid phase separation in the cell. In this short review, we show that such non-stoichiometric organization may derive from the heterogeneity of the system, a mismatch in valency and/or geometry of the partners, and/or intrinsic structural disorder and multivalency of the component proteins.
    [Show full text]