Germline P Granules Are Liquid Droplets That Localize by Controlled Dissolution/Condensation

doi:10.1126/SCIENCE.1172046

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Germline P Granules Are Liquid Droplets That Localize by Controlled Dissolution/Condensation

Journal Article•DOI•

Germline P Granules Are Liquid Droplets That Localize by Controlled Dissolution/Condensation

Clifford P. Brangwynne¹, Christian R. Eckmann¹, David S. Courson², Agata Rybarska¹, Carsten Hoege¹, Jöbin Gharakhani¹, Jöbin Gharakhani², Frank Jülicher¹, Frank Jülicher², Anthony A. Hyman², Anthony A. Hyman¹ - Show less +7 more•Institutions (2)

Max Planck Society¹, Marine Biological Laboratory²

26 Jun 2009-Science (American Association for the Advancement of Science)-Vol. 324, Iss: 5935, pp 1729-1732

TL;DR: It is shown that P granules exhibit liquid-like behaviors, including fusion, dripping, and wetting, which is used to estimate their viscosity and surface tension, and reflects a classic phase transition, in which polarity proteins vary the condensation point across the cell.

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Abstract: In sexually reproducing organisms, embryos specify germ cells, which ultimately generate sperm and eggs In Caenorhabditis elegans, the first germ cell is established when RNA and protein-rich P granules localize to the posterior of the one-cell embryo Localization of P granules and their physical nature remain poorly understood Here we show that P granules exhibit liquid-like behaviors, including fusion, dripping, and wetting, which we used to estimate their viscosity and surface tension As with other liquids, P granules rapidly dissolved and condensed Localization occurred by a biased increase in P granule condensation at the posterior This process reflects a classic phase transition, in which polarity proteins vary the condensation point across the cell Such phase transitions may represent a fundamental physicochemical mechanism for structuring the cytoplasm

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Journal Article•DOI•

Biomolecular condensates: organizers of cellular biochemistry

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Salman F. Banani¹, Hyun-Ok Kate Lee², Anthony A. Hyman², Michael K. Rosen¹•Institutions (2)

University of Texas Southwestern Medical Center¹, Max Planck Society²

01 May 2017-Nature Reviews Molecular Cell Biology

TL;DR: This work has shown that liquid–liquid phase separation driven by multivalent macromolecular interactions is an important organizing principle for biomolecular condensates and has proposed a physical framework for this organizing principle.

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Abstract: In addition to membrane-bound organelles, eukaryotic cells feature various membraneless compartments, including the centrosome, the nucleolus and various granules. Many of these compartments form through liquid–liquid phase separation, and the principles, mechanisms and regulation of their assembly as well as their cellular functions are now beginning to emerge. Biomolecular condensates are micron-scale compartments in eukaryotic cells that lack surrounding membranes but function to concentrate proteins and nucleic acids. These condensates are involved in diverse processes, including RNA metabolism, ribosome biogenesis, the DNA damage response and signal transduction. Recent studies have shown that liquid–liquid phase separation driven by multivalent macromolecular interactions is an important organizing principle for biomolecular condensates. With this physical framework, it is now possible to explain how the assembly, composition, physical properties and biochemical and cellular functions of these important structures are regulated.

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3,294 citations

Journal Article•DOI•

Liquid phase condensation in cell physiology and disease.

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Yongdae Shin¹, Clifford P. Brangwynne¹•Institutions (1)

Princeton University¹

22 Sep 2017-Science

TL;DR: The findings together suggest that several membrane-less organelles have been shown to exhibit a concentration threshold for assembly, a hallmark of phase separation, and represent liquid-phase condensates, which form via a biologically regulated (liquid-liquid) phase separation process.

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Abstract: BACKGROUND Living cells contain distinct subcompartments to facilitate spatiotemporal regulation of biological reactions. In addition to canonical membrane-bound organelles such as secretory vesicles and endoplasmic reticulum, there are many organelles that do not have an enclosing membrane yet remain coherent structures that can compartmentalize and concentrate specific sets of molecules. Examples include assemblies in the nucleus such as the nucleolus, Cajal bodies, and nuclear speckles and also cytoplasmic structures such as stress granules, P-bodies, and germ granules. These structures play diverse roles in various biological processes and are also increasingly implicated in protein aggregation diseases. ADVANCES A number of studies have shown that membrane-less assemblies exhibit remarkable liquid-like features. As with conventional liquids, they typically adopt round morphologies and coalesce into a single droplet upon contact with one another and also wet intracellular surfaces such as the nuclear envelope. Moreover, component molecules exhibit dynamic exchange with the surrounding nucleoplasm and cytoplasm. These findings together suggest that these structures represent liquid-phase condensates, which form via a biologically regulated (liquid-liquid) phase separation process. Liquid phase condensation increasingly appears to be a fundamental mechanism for organizing intracellular space. Consistent with this concept, several membrane-less organelles have been shown to exhibit a concentration threshold for assembly, a hallmark of phase separation. At the molecular level, weak, transient interactions between molecules with multivalent domains or intrinsically disordered regions (IDRs) are a driving force for phase separation. In cells, condensation of liquid-phase assemblies can be regulated by active processes, including transcription and various posttranslational modifications. The simplest physical picture of a homogeneous liquid phase is often not enough to capture the full complexity of intracellular condensates, which frequently exhibit heterogeneous multilayered structures with partially solid-like characters. However, recent studies have shown that multiple distinct liquid phases can coexist and give rise to richly structured droplet architectures determined by the relative liquid surface tensions. Moreover, solid-like phases can emerge from metastable liquid condensates via multiple routes of potentially both kinetic and thermodynamic origins, which has important implications for the role of intracellular liquids in protein aggregation pathologies. OUTLOOK The list of intracellular assemblies driven by liquid phase condensation is growing rapidly, but our understanding of their sequence-encoded biological function and dysfunction lags behind. Moreover, unlike equilibrium phases of nonliving matter, living cells are far from equilibrium, with intracellular condensates subject to various posttranslational regulation and other adenosine triphosphate–dependent biological activity. Efforts using in vitro reconstitution, combined with traditional cell biology approaches and quantitative biophysical tools, are required to elucidate how such nonequilibrium features of living cells control intracellular phase behavior. The functional consequences of forming liquid condensates are likely multifaceted and may include facilitated reaction, sequestration of specific factors, and organization of associated intracellular structures. Liquid phase condensation is particularly interesting in the nucleus, given the growing interest in the impact of nuclear phase behavior on the flow of genetic information; nuclear condensates range from micrometer-sized bodies such as the nucleolus to submicrometer structures such as transcriptional assemblies, all of which directly interact with and regulate the genome. Deepening our understanding of these intracellular states of matter not only will shed light on the basic biology of cellular organization but also may enable therapeutic intervention in protein aggregation disease by targeting intracellular phase behavior.

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2,432 citations

Cites background from "Germline P Granules Are Liquid Drop..."

...drip off the surface of a nucleus in response to shear (12)....
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...The list of condensates thought to assemble through liquid-liquid phase separation now includes many RNP bodies such as P granules (12), nucleoli (13), various signaling clusters (17, 136), and numerous other related structures....
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...liquid phase nature of P granules (12), nucleoli (13, 64, 65), and dozens of other such fluid...
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Journal Article•DOI•

Liquid-liquid phase separation in biology.

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Anthony A. Hyman¹, Christoph Weber¹, Frank Jülicher¹•Institutions (1)

Max Planck Society¹

06 Oct 2014-Annual Review of Cell and Developmental Biology

TL;DR: The basic physical concepts necessary to understand the consequences of liquid-like states for biological functions are discussed.

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Abstract: Cells organize many of their biochemical reactions in non-membrane compartments. Recent evidence has shown that many of these compartments are liquids that form by phase separation from the cytoplasm. Here we discuss the basic physical concepts necessary to understand the consequences of liquid-like states for biological functions.

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2,088 citations

Cites background from "Germline P Granules Are Liquid Drop..."

...Careful observation showed that they fuse, exchange components rapidly with the cytoplasm, are easily deformed by flows, and have a viscosity similar to runny honey (Brangwynne et al. 2009)....
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...(a,c,d ) Modified with permission from Brangwynne et al. (2009)....
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...Therefore, although nucleoli are considerably more viscous than P granules, they both have liquid-like properties (Brangwynne et al. 2009, 2011)....
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...39 CB30CH03-Hyman ARI 11 September 2014 7:1...
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...Initially they were called granules because of their particulate appearance, but closer inspection of their dynamics (Brangwynne et al. 2009) reveals that they are better described as liquids for the following reasons: 1....
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Journal Article•DOI•

A Liquid-to-Solid Phase Transition of the ALS Protein FUS Accelerated by Disease Mutation

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Avinash Patel¹, Hyun O. Lee¹, Louise Jawerth¹, Shovamayee Maharana¹, Marcus Jahnel¹, Marco Y. Hein¹, Stoyno S. Stoynov², Julia Mahamid¹, Shambaditya Saha¹, Titus M. Franzmann¹, Andrej Pozniakovski¹, Ina Poser¹, Nicola Maghelli¹, Loic Royer¹, Martin Weigert¹, Eugene W. Myers¹, Stephan W. Grill¹, David N. Drechsel¹, Anthony A. Hyman¹, Simon Alberti¹ - Show less +16 more•Institutions (2)

Max Planck Society¹, Bulgarian Academy of Sciences²

27 Aug 2015-Cell

TL;DR: It is proposed that liquid-like compartments carry the trade-off between functionality and risk of aggregation and that aberrant phase transitions within liquid- like compartments lie at the heart of ALS and, presumably, other age-related diseases.

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1,988 citations

Cites background or methods from "Germline P Granules Are Liquid Drop..."

...Examples include germ (P) granules (Brangwynne et al., 2009), processing (P/GW)...
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...Using the relaxation time and the FRAP (fluorescence recovery after photobleaching) times, we approximated viscosity values, as previously described (Brangwynne et al., 2009)....
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...Recently, an increasing number of these non-membrane-bound compartments have been shown to behave like condensed liquid phases of the cytoplasm or nucleoplasm (Aggarwal et al., 2013; Brangwynne et al., 2009, 2011; Hubstenberger et al., 2013; Lee et al., 2013; Wippich et al., 2013)....
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...Examples include germ (P) granules (Brangwynne et al., 2009), processing (P/GW) 1066 Cell 162, 1066–1077, August 27, 2015 ª2015 Elsevier Inc. bodies (Kedersha et al., 2005), stress granules (Wippich et al., 2013), nucleoli (Brangwynne et al., 2011), Cajal bodies (Strzelecka et al., 2010) and likely…...
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...The constant mixing of components within a liquid can be tested by a technique known as ‘‘half-bleach’’ (Brangwynne et al., 2009; Hyman et al., 2014)....
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Journal Article•DOI•

Phase Separation by Low Complexity Domains Promotes Stress Granule Assembly and Drives Pathological Fibrillization

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Amandine Molliex¹, Jamshid Temirov¹, Jihun Lee¹, Maura Coughlin¹, Anderson P. Kanagaraj¹, Hong Joo Kim¹, Tanja Mittag¹, J. Paul Taylor¹ - Show less +4 more•Institutions (1)

St. Jude Children's Research Hospital¹

24 Sep 2015-Cell

TL;DR: It is demonstrated that the disease-related RBP hnRNPA1 undergoes liquid-liquid phase separation (LLPS) into protein-rich droplets mediated by a low complexity sequence domain (LCD), and suggested that LCD-mediated LLPS contributes to the assembly of stress granules and their liquid properties.

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1,947 citations

Cites background from "Germline P Granules Are Liquid Drop..."

...LLPS has been proposed as the molecular mechanism underlying formation of membrane-less cellular bodies that exhibit liquid properties, such as P granules and nucleoli (Brangwynne et al., 2009; Elbaum-Garfinkle et al., 2015; Fromm et al., 2014; Li et al., 2012; Nott et al., 2015)....
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...It has recently emerged that cells organize many biochemical processes in membrane-less compartments that have liquidlike properties, exemplified by germ granules in C. elegans and nucleoli in X. laevis (Brangwynne et al., 2009, 2011)....
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PGL-1, a Predicted RNA-Binding Component of Germ Granules, Is Essential for Fertility in C. elegans

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Indiana University¹, University of Colorado Boulder²

04 Sep 1998-Cell

TL;DR: It is shown that a component of germ granules is essential for fertility in C. elegans and that its primary function is in germline proliferation.

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