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  1. Article ; Online: Macromolecular crowding: Sensing without a sensor.

    Holt, Liam J / Delarue, Morgan

    Current opinion in cell biology

    2023  Volume 85, Page(s) 102269

    Abstract: All living cells are crowded with macromolecules. Crowding can directly modulate biochemical reactions to various degrees depending on the sizes, shapes, and binding affinities of the reactants. Here, we explore the possibility that cells can sense and ... ...

    Abstract All living cells are crowded with macromolecules. Crowding can directly modulate biochemical reactions to various degrees depending on the sizes, shapes, and binding affinities of the reactants. Here, we explore the possibility that cells can sense and adapt to changes in crowding through the widespread modulation of biochemical reactions without the need for a dedicated sensor. Additionally, we explore phase separation as a general physicochemical response to changes in crowding, and a mechanism to both transduce information and physically restore crowding homeostasis.
    MeSH term(s) Macromolecular Substances/metabolism ; Cell Physiological Phenomena
    Chemical Substances Macromolecular Substances
    Language English
    Publishing date 2023-10-26
    Publishing country England
    Document type Journal Article ; Review
    ZDB-ID 1026381-0
    ISSN 1879-0410 ; 0955-0674
    ISSN (online) 1879-0410
    ISSN 0955-0674
    DOI 10.1016/j.ceb.2023.102269
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Increased mesoscale diffusivity in response to acute glucose starvation.

    Xie, Ying / Gresham, David / Holt, Liam J

    microPublication biology

    2023  Volume 2023

    Abstract: Macromolecular crowding is an important property of cells that impacts multiple biological processes. Passive microrheology using single particle tracking is a powerful means of studying macromolecular crowding. Here we monitored the diffusivity of self- ... ...

    Abstract Macromolecular crowding is an important property of cells that impacts multiple biological processes. Passive microrheology using single particle tracking is a powerful means of studying macromolecular crowding. Here we monitored the diffusivity of self-assembling fluorescent nanoparticles (μNS) and mRNPs (
    Language English
    Publishing date 2023-02-21
    Publishing country United States
    Document type Journal Article
    ISSN 2578-9430
    ISSN (online) 2578-9430
    DOI 10.17912/micropub.biology.000729
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: How it feels in a cell.

    Bonucci, Martina / Shu, Tong / Holt, Liam J

    Trends in cell biology

    2023  Volume 33, Issue 11, Page(s) 924–938

    Abstract: Life emerges from thousands of biochemical processes occurring within a shared intracellular environment. We have gained deep insights from in vitro reconstitution of isolated biochemical reactions. However, the reaction medium in test tubes is typically ...

    Abstract Life emerges from thousands of biochemical processes occurring within a shared intracellular environment. We have gained deep insights from in vitro reconstitution of isolated biochemical reactions. However, the reaction medium in test tubes is typically simple and diluted. The cell interior is far more complex: macromolecules occupy more than a third of the space, and energy-consuming processes agitate the cell interior. Here, we review how this crowded, active environment impacts the motion and assembly of macromolecules, with an emphasis on mesoscale particles (10-1000 nm diameter). We describe methods to probe and analyze the biophysical properties of cells and highlight how changes in these properties can impact physiology and signaling, and potentially contribute to aging, and diseases, including cancer and neurodegeneration.
    Language English
    Publishing date 2023-06-05
    Publishing country England
    Document type Journal Article ; Review
    ZDB-ID 30122-x
    ISSN 1879-3088 ; 0962-8924
    ISSN (online) 1879-3088
    ISSN 0962-8924
    DOI 10.1016/j.tcb.2023.05.002
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  4. Article: mRNA condensation fluidizes the cytoplasm.

    Xie, Ying / Liu, Tiewei / Gresham, David / Holt, Liam J

    bioRxiv : the preprint server for biology

    2023  

    Abstract: The intracellular environment is packed with macromolecules of mesoscale size, and this crowded milieu significantly influences cell physiology. When exposed to stress, mRNAs released after translational arrest condense with RNA binding proteins, ... ...

    Abstract The intracellular environment is packed with macromolecules of mesoscale size, and this crowded milieu significantly influences cell physiology. When exposed to stress, mRNAs released after translational arrest condense with RNA binding proteins, resulting in the formation of membraneless RNA protein (RNP) condensates known as processing bodies (P-bodies) and stress granules (SGs). However, the impact of the assembly of these condensates on the biophysical properties of the crowded cytoplasmic environment remains unclear. Here, we find that upon exposure to stress, polysome collapse and condensation of mRNAs increases mesoscale particle diffusivity in the cytoplasm. Increased mesoscale diffusivity is required for the efficient formation of Q-bodies, membraneless organelles that coordinate degradation of misfolded peptides that accumulate during stress. Additionally, we demonstrate that polysome collapse and stress granule formation has a similar effect in mammalian cells, fluidizing the cytoplasm at the mesoscale. We find that synthetic, light-induced RNA condensation is sufficient to fluidize the cytoplasm, demonstrating a causal effect of RNA condensation. Together, our work reveals a new functional role for stress-induced translation inhibition and formation of RNP condensates in modulating the physical properties of the cytoplasm to effectively respond to stressful conditions.
    Language English
    Publishing date 2023-07-15
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2023.05.30.542963
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Condensation of LINE-1 is critical for retrotransposition.

    Sil, Srinjoy / Keegan, Sarah / Ettefa, Farida / Denes, Lance T / Boeke, Jef D / Holt, Liam J

    eLife

    2023  Volume 12

    Abstract: LINE-1 (L1) is the only autonomously active retrotransposon in the human genome, and accounts for 17% of the human genome. The L1 mRNA encodes two proteins, ORF1p and ORF2p, both essential for retrotransposition. ORF2p has reverse transcriptase and ... ...

    Abstract LINE-1 (L1) is the only autonomously active retrotransposon in the human genome, and accounts for 17% of the human genome. The L1 mRNA encodes two proteins, ORF1p and ORF2p, both essential for retrotransposition. ORF2p has reverse transcriptase and endonuclease activities, while ORF1p is a homotrimeric RNA-binding protein with poorly understood function. Here, we show that condensation of ORF1p is critical for L1 retrotransposition. Using a combination of biochemical reconstitution and live-cell imaging, we demonstrate that electrostatic interactions and trimer conformational dynamics together tune the properties of ORF1p assemblies to allow for efficient L1 ribonucleoprotein (RNP) complex formation in cells. Furthermore, we relate the dynamics of ORF1p assembly and RNP condensate material properties to the ability to complete the entire retrotransposon life-cycle. Mutations that prevented ORF1p condensation led to loss of retrotransposition activity, while orthogonal restoration of coiled-coil conformational flexibility rescued both condensation and retrotransposition. Based on these observations, we propose that dynamic ORF1p oligomerization on L1 RNA drives the formation of an L1 RNP condensate that is essential for retrotransposition.
    MeSH term(s) Humans ; Retroelements/genetics ; Long Interspersed Nucleotide Elements/genetics ; Mutation ; RNA-Binding Proteins/metabolism ; RNA/metabolism
    Chemical Substances Retroelements ; RNA-Binding Proteins ; RNA (63231-63-0)
    Language English
    Publishing date 2023-04-28
    Publishing country England
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2687154-3
    ISSN 2050-084X ; 2050-084X
    ISSN (online) 2050-084X
    ISSN 2050-084X
    DOI 10.7554/eLife.82991
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  6. Article ; Online: Epithelial tissue confinement inhibits cell growth and leads to volume-reducing divisions.

    Devany, John / Falk, Martin J / Holt, Liam J / Murugan, Arvind / Gardel, Margaret L

    Developmental cell

    2023  Volume 58, Issue 16, Page(s) 1462–1476.e8

    Abstract: Cell proliferation is a central process in tissue development, homeostasis, and disease, yet how proliferation is regulated in the tissue context remains poorly understood. Here, we introduce a quantitative framework to elucidate how tissue growth ... ...

    Abstract Cell proliferation is a central process in tissue development, homeostasis, and disease, yet how proliferation is regulated in the tissue context remains poorly understood. Here, we introduce a quantitative framework to elucidate how tissue growth dynamics regulate cell proliferation. Using MDCK epithelial monolayers, we show that a limiting rate of tissue expansion creates confinement that suppresses cell growth; however, this confinement does not directly affect the cell cycle. This leads to uncoupling between rates of cell growth and division in epithelia and, thereby, reduces cell volume. Division becomes arrested at a minimal cell volume, which is consistent across diverse epithelia in vivo. Here, the nucleus approaches the minimum volume capable of packaging the genome. Loss of cyclin D1-dependent cell-volume regulation results in an abnormally high nuclear-to-cytoplasmic volume ratio and DNA damage. Overall, we demonstrate how epithelial proliferation is regulated by the interplay between tissue confinement and cell-volume regulation.
    MeSH term(s) Epithelial Cells/metabolism ; Cell Cycle/physiology ; Cell Division ; Epithelium ; Cell Proliferation
    Language English
    Publishing date 2023-06-19
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2054967-2
    ISSN 1878-1551 ; 1534-5807
    ISSN (online) 1878-1551
    ISSN 1534-5807
    DOI 10.1016/j.devcel.2023.05.018
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  7. Article ; Online: Publisher Correction: Reciprocal regulation of cellular mechanics and metabolism.

    Evers, Tom M J / Holt, Liam J / Alberti, Simon / Mashaghi, Alireza

    Nature metabolism

    2021  Volume 3, Issue 6, Page(s) 876–877

    Language English
    Publishing date 2021-06-09
    Publishing country Germany
    Document type Published Erratum
    ISSN 2522-5812
    ISSN (online) 2522-5812
    DOI 10.1038/s42255-021-00423-6
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  8. Article ; Online: Reciprocal regulation of cellular mechanics and metabolism.

    Evers, Tom M J / Holt, Liam J / Alberti, Simon / Mashaghi, Alireza

    Nature metabolism

    2021  Volume 3, Issue 4, Page(s) 456–468

    Abstract: Metabolism and mechanics are intrinsically intertwined. External forces, sensed through the cytoskeleton or distortion of the cell and organelles, induce metabolic changes in the cell. The resulting changes in metabolism, in turn, feed back to regulate ... ...

    Abstract Metabolism and mechanics are intrinsically intertwined. External forces, sensed through the cytoskeleton or distortion of the cell and organelles, induce metabolic changes in the cell. The resulting changes in metabolism, in turn, feed back to regulate every level of cell biology, including the mechanical properties of cells and tissues. Here we examine the links between metabolism and mechanics, highlighting signalling pathways involved in the regulation and response to cellular mechanosensing. We consider how forces and metabolism regulate one another through nanoscale molecular sensors, micrometre-scale cytoskeletal networks, organelles and dynamic biomolecular condensates. Understanding this cross-talk will create diagnostic and therapeutic opportunities for metabolic disorders such as cancer, cardiovascular pathologies and obesity.
    MeSH term(s) Animals ; Cell Biology ; Cytoskeleton/physiology ; Humans ; Mechanotransduction, Cellular/physiology ; Metabolism/physiology ; Signal Transduction ; Stress, Mechanical
    Language English
    Publishing date 2021-04-19
    Publishing country Germany
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
    ISSN 2522-5812
    ISSN (online) 2522-5812
    DOI 10.1038/s42255-021-00384-w
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  9. Article ; Online: Control of nuclear size by osmotic forces in

    Lemière, Joël / Real-Calderon, Paula / Holt, Liam J / Fai, Thomas G / Chang, Fred

    eLife

    2022  Volume 11

    Abstract: The size of the nucleus scales robustly with cell size so that the nuclear-to-cell volume ratio (N/C ratio) is maintained during cell growth in many cell types. The mechanism responsible for this scaling remains mysterious. Previous studies have ... ...

    Abstract The size of the nucleus scales robustly with cell size so that the nuclear-to-cell volume ratio (N/C ratio) is maintained during cell growth in many cell types. The mechanism responsible for this scaling remains mysterious. Previous studies have established that the N/C ratio is not determined by DNA amount but is instead influenced by factors such as nuclear envelope mechanics and nuclear transport. Here, we developed a quantitative model for nuclear size control based upon colloid osmotic pressure and tested key predictions in the fission yeast
    MeSH term(s) Active Transport, Cell Nucleus/physiology ; Cell Nucleus/metabolism ; Nuclear Envelope/metabolism ; Schizosaccharomyces/genetics ; Schizosaccharomyces pombe Proteins/metabolism
    Chemical Substances Schizosaccharomyces pombe Proteins
    Language English
    Publishing date 2022-07-20
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, Non-U.S. Gov't
    ZDB-ID 2687154-3
    ISSN 2050-084X ; 2050-084X
    ISSN (online) 2050-084X
    ISSN 2050-084X
    DOI 10.7554/eLife.76075
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  10. Article: Macromolecular crowding limits growth under pressure.

    Alric, Baptiste / Formosa-Dague, Cécile / Dague, Etienne / Holt, Liam J / Delarue, Morgan

    Nature physics

    2022  Volume 18, Issue 4, Page(s) 411–416

    Abstract: Cells that grow in confined spaces eventually build up mechanical compressive stress. This growth-induced pressure (GIP) decreases cell growth. GIP is important in a multitude of contexts from cancer, to microbial infections, to biofouling, yet our ... ...

    Abstract Cells that grow in confined spaces eventually build up mechanical compressive stress. This growth-induced pressure (GIP) decreases cell growth. GIP is important in a multitude of contexts from cancer, to microbial infections, to biofouling, yet our understanding of its origin and molecular consequences remains limited. Here, we combine microfluidic confinement of the yeast
    Language English
    Publishing date 2022-02-24
    Publishing country England
    Document type Journal Article
    ZDB-ID 2206346-8
    ISSN 1745-2481 ; 1745-2473
    ISSN (online) 1745-2481
    ISSN 1745-2473
    DOI 10.1038/s41567-022-01506-1
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