UMR168 – Physico-Chimie Curie Lab

Team Publications

Year of publication 2019

Attner MA*, Keil W*, Benavidez JM, Greenwald I (2019 Sep 23)

HLH-2/E2A Expression Links Stochastic and Deterministic Elements of a Cell Fate Decision during C. elegans Gonadogenesis

Current Biology : 29 : 1-7 : DOI : https://doi.org/10.1016/j.cub.2019.07.062 Learn more
Summary

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Mathieu Richard, Carles Blanch-Mercader, Hajer Ennomani, Wenxiang Cao, Enrique M De La Cruz, Jean-François Joanny, Frank Jülicher, Laurent Blanchoin, Pascal Martin (2019 Jul 11)

Active cargo positioning in antiparallel transport networks.

Proceedings of the National Academy of Sciences of the United States of America : DOI : 10.1073/pnas.1900416116 Learn more
Summary

Cytoskeletal filaments assemble into dense parallel, antiparallel, or disordered networks, providing a complex environment for active cargo transport and positioning by molecular motors. The interplay between the network architecture and intrinsic motor properties clearly affects transport properties but remains poorly understood. Here, by using surface micropatterns of actin polymerization, we investigate stochastic transport properties of colloidal beads in antiparallel networks of overlapping actin filaments. We found that 200-nm beads coated with myosin Va motors displayed directed movements toward positions where the net polarity of the actin network vanished, accumulating there. The bead distribution was dictated by the spatial profiles of local bead velocity and diffusion coefficient, indicating that a diffusion-drift process was at work. Remarkably, beads coated with heavy-mero-myosin II motors showed a similar behavior. However, although velocity gradients were steeper with myosin II, the much larger bead diffusion observed with this motor resulted in less precise positioning. Our observations are well described by a 3-state model, in which active beads locally sense the net polarity of the network by frequently detaching from and reattaching to the filaments. A stochastic sequence of processive runs and diffusive searches results in a biased random walk. The precision of bead positioning is set by the gradient of net actin polarity in the network and by the run length of the cargo in an attached state. Our results unveiled physical rules for cargo transport and positioning in networks of mixed polarity.

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Katz M, Corson F*, Keil W*, Singhal A, Bae A, Lu Y, Liang Y & Shaham S (2019 Apr 23)

Glutamate spillover in C. elegans triggers repetitive behavior through presynaptic activation of MGL-2/mGluR5

Nature Communications : 10 : DOI : 10.1038/s41467-019-09581-4 Learn more
Summary

Glutamate is a major excitatory neurotransmitter, and impaired glutamate clearance following synaptic release promotes spillover, inducing extra-synaptic signaling. The effects of glutamate spillover on animal behavior and its neural correlates are poorly understood. We developed a glutamate spillover model in Caenorhabditis elegans by inactivating the conserved glial glutamate transporter GLT-1. GLT-1 loss drives aberrant repetitive locomotory reversal behavior through uncontrolled oscillatory release of glutamate onto AVA, a major interneuron governing reversals. Repetitive glutamate release and reversal behavior require the glutamate receptor MGL-2/mGluR5, expressed in RIM and other interneurons presynaptic to AVA. mgl-2 loss blocks oscillations and repetitive behavior; while RIM activation is sufficient to induce repetitive reversals in glt-1 mutants. Repetitive AVA firing and reversals require EGL-30/Gαq, an mGluR5 effector. Our studies reveal that cyclic autocrine presynaptic activation drives repetitive reversals following glutamate spillover. That mammalian GLT1 and mGluR5 are implicated in pathological motor repetition suggests a common mechanism controlling repetitive behaviors.

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Mélanie Tobin, Atitheb Chaiyasitdhi, Vincent Michel, Nicolas Michalski, Pascal Martin (2019 Apr 2)

Stiffness and tension gradients of the hair cell’s tip-link complex in the mammalian cochlea.

eLife : DOI : 10.7554/eLife.43473 Learn more
Summary

Sound analysis by the cochlea relies on frequency tuning of mechanosensory hair cells along a tonotopic axis. To clarify the underlying biophysical mechanism, we have investigated the micromechanical properties of the hair cell’s mechanoreceptive hair bundle within the apical half of the rat cochlea. We studied both inner and outer hair cells, which send nervous signals to the brain and amplify cochlear vibrations, respectively. We find that tonotopy is associated with gradients of stiffness and resting mechanical tension, with steeper gradients for outer hair cells, emphasizing the division of labor between the two hair-cell types. We demonstrate that tension in the tip links that convey force to the mechano-electrical transduction channels increases at reduced Ca. Finally, we reveal gradients in stiffness and tension at the level of a single tip link. We conclude that mechanical gradients of the tip-link complex may help specify the characteristic frequency of the hair cell.

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Jamecna D, Polidori DJ, Mesmin B, Dezi M, Lévy D, Bigay J, Antonny B (2019 Mar 22)

An intrinsically disordered region in OSBP acts as an entropic barrier to control protein dynamics and orientation at membrane contact sites

Developmental cell * : * highlighted Trend in Cell Biology 2019 : DOI : 10.1016/j.devcel.2019.02.021 Learn more
Summary

Lipid transfer proteins (LTPs) acting at membrane contact sites (MCS) between the ER and other organelles contain domains involved in heterotypic (e.g. ER to Golgi) membrane tethering as well as domains involved in lipid transfer. Here, we show that a long ≈ 90 aa intrinsically unfolded sequence at the N-terminus of oxysterol binding protein (OSBP) controls OSBP orientation and dynamics at MCS. This Gly-Pro-Ala-rich sequence, whose hydrodynamic radius is twice as that of folded domains, prevents the two PH domains of the OSBP dimer from homotypically tethering two Golgi-like membranes and considerably facilitates OSBP in-plane diffusion and recycling at MCS. Although quite distant in sequence, the N-terminus of OSBP-related protein-4 (ORP4) has similar effects. We propose that N-terminal sequences of low complexity in ORPs form an entropic barrier that restrains protein orientation, limits protein density and facilitates protein mobility in the narrow and crowded MCS environment.

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Simon C*, Kusters R*, Caorsi V*, Allard A, Abou-Ghali M, Manzi J, Di Cicco A, Lévy D, Lenz M, Joanny J-F, Campillo C, Plastino J, Sens P*, Sykes C* (2019 Mar 18)

Actin dynamics drive cell-like membrane deformation

Nature Physics : DOI : 10.1038/s41567-019-0464-1 Learn more
Summary

Cell membrane deformations are crucial for proper cell function. Specialized protein assemblies initiate inward or outward membrane deformations that the cell uses respectively to uptake external substances or probe the environment. The assembly and dynamics of the actin cytoskeleton are involved in this process, although their detailed role remains controversial. We show here that a dynamic, branched actin network is sufficient to initiate both inward and outward membrane deformation. The polymerization of a dense actin network at the membrane of liposomes produces inward membrane bending at low tension, while outward deformations are robustly generated regardless of tension. Our results shed light on the mechanism cells use to internalize material, both in mammalian cells, where actin polymerization forces are required when membrane tension is increased, and in yeast, where those forces are necessary to overcome the opposing turgor pressure. By combining experimental observations with physical modelling, we propose a mechanism that explains how membrane tension and the architecture of the actin network regulate cell-like membrane deformations.

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Bertin Aurélie, Lomakin Alexis (2019 Feb 15)

Meeting report – Building the Cell 2018

journal of cell science : 132 : DOI : 10.1242/jcs.229765 Learn more
Summary

Cell biologists from all around the world gathered in Paris on the 26 to 28 September 2018 to participate in the 3rd international meeting ‘Building the Cell’. It was organized by Hélène Barelli, Arnaud Echard, Thierry Galli, Florence Niedergang, Manuel Théry and Marie Hélène Verlhac on behalf of the French Society for Cell Biology (SBCF) at the Institut Pasteur. Around 230 participants joined the meeting for stimulating talks, discussions, poster sessions, and a gala dinner on the Seine that included a music performance by the rock group ‘Membrane Band’. The unifying theme of the meeting was the development of creative multidisciplinary approaches to understand cellular life at different scales in a dynamic and quantitative manner. Here, we summarize the results presented at the meeting and the emerging ideas from the different sessions.

The 3rd international meeting ‘Building the Cell’ (Fig. 1Fig. 2) was divided into ten different sessions that covered a variety of topics including intracellular trafficking, cell division, cytoskeletal dynamics and cell mechanics, cancer and stem cell biology, embryonic development and tissue morphogenesis, neurobiology, and aggregates and phase transitions. A broad spectrum of modern approaches and experimental systems ranging from synthetic biology and stem cell technologies to 3D organoids and animal models was presented. The meeting highlighted some of the latest and novel findings in cell biology, often coupled to major methodological developments in quantitative microscopy and computational modelling, as well as cell and tissue micro-engineering.

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Sarah Moitrier, Carles Blanch-Mercader, Simon Garcia, Kristina Sliogeryte,abc Tobias Martin, Jacques Camonis, Philippe Marcq, Pascal Silberzan and Isabelle Bonnet (2019 Feb 4)

Collective stresses drive competition between monolayers of normal and Ras-transformed cells

Soft Matter : 15 : DOI : 10.1039/C8SM01523F Learn more
Summary

We study the competition for space between two cell lines that differ only in the expression of the Ras oncogene. The two cell populations are initially separated and set to migrate antagonistically towards an in-between stripe of free substrate. After contact, their interface moves towards the population of normal cells. We interpret the velocity and traction force data taken before and after contact thanks to a hydrodynamic description of collectively migrating cohesive cell sheets. The kinematics of cells, before and after contact, allows us to estimate the relative material parameters for both cell lines. As predicted by the model, the transformed cell population with larger collective stresses pushes the wild type cell population.

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Dreier J, Castello M, Coceano G, Cáceres R, Plastino J, Vicidomini G, Testa I (2019 Feb 1)

Smart scanning for low-illumination and fast RESOLFT nanoscopy in vivo

Nature Communications : 10 : 556 : DOI : 10.1038/s41467-019-08442-4 Learn more
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Beber A, Taveneau C, Nania M, Tsai FC, Di Cicco A, Bassereau P, Lévy D, Cabral JT, Isambert H, Mangenot S*, Bertin A* (2019 Jan 24)

Membrane reshaping by micrometric curvature sensitive septin filaments

Nature communications : DOI : 10.1038/s41467-019-08344-5 Learn more
Summary

Septins are cytoskeletal filaments that assemble at the inner face of the plasma membrane.They are localized at constriction sites and impact membrane remodeling. We report in vitro tools to examine how yeast septins behave on curved and deformable membranes. Septins reshape the membranes of Giant Unilamellar Vesicles with the formation of periodic spikes, while flattening smaller vesicles. We show that membrane deformations are associated to preferential arrangement of Septin filaments on specific curvatures. When binding to bilayers supported on custom-designed periodic wavy patterns displaying positive and negative micrometric radii of curvatures, septin filaments remain straight and perpendicular to the curvature of the convex parts, while bending negatively to follow concave geometries. Based on these results, we propose a theoretical model that describes the deformations and micrometric curvature sensitivity observed in vitro. The model captures the reorganizations of septin filaments throughout cytokinesis in vivo, providing mechanistic insights into cell division.

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Kelley LC, Chi Q, Cáceres R, Hastie E, Schindler AJ, Jiang Y, Matus DQ, Plastino J, Sherwood DR (2019 Jan 24)

Adaptive F-actin polymerization and localized ATP production drive basement membrane invasion in the absence of MMPs

Developmental Cell : 48 : 313-328 : DOI : 10.1016/j.devcel.2018.12.018 Learn more
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Year of publication 2018

McHugh T, Zou J, Volkov VA, Bertin A, Rappsilber J, Dogterom M, Welburn JPI (2018 Dec 21)

The depolymerase activity of MCAK shows graded response to Aurora B kinase phosphorylation through allosteric regulation.

journal of cell science : DOI : 10.1242/jcs.228353 Learn more
Summary

Kinesin-13 motors regulate precise microtubule dynamics and limit microtubule length throughout metazoans by depolymerizing microtubule ends. Recently, MCAK has been proposed to undergo large conformational changes during its catalytic cycle, as it switches from solution to bound state. Here, we reveal that MCAK has a compact conformation in solution using crosslinking and electron microscopy. When MCAK is bound to the microtubule ends, it adopts an extended conformation with the N terminus and neck region of MCAK interacting with the microtubule. Interestingly, the region of MCAK that interacts with the microtubule is the region phosphorylated by Aurora B and contains an EB-binding motif. The level of phosphorylation of the N terminus results in a graded microtubule depolymerase activity. Here we show the N terminus of MCAK forms a platform to integrate Aurora B kinase downstream signals and in response fine-tunes its depolymerase activity during mitosis. We propose that this allosteric control mechanism allows decoupling of the N terminus from the motor domain of MCAK to allow MCAK depolymerase activity at kinetochores.

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Kutscher LM, Keil W, Shaham S (2018 Oct 22)

RAB-35 and ARF-6 GTPases Mediate Engulfment and Clearance Following Linker Cell-Type Death

Developmental Cell : 47 : 222-238 : DOI : 10.1016/j.devcel.2018.08.015 Learn more
Summary

Clearance of dying cells is essential for development and homeostasis. Conserved genes mediate apoptotic cell removal, but whether these genes control non-apoptotic cell removal is a major open question. Linker cell-type death (LCD) is a prevalent non-apoptotic developmental cell death process with features conserved from C. elegans to vertebrates. Using microfluidics-based long-term in vivo imaging, we show that unlike apoptotic cells, the C. elegans linker cell, which dies by LCD, is competitively phagocytosed by two neighboring cells, resulting in cell splitting. Subsequent cell elimination does not require apoptotic engulfment genes. Rather, we find that RAB-35 GTPase is a key coordinator of competitive phagocytosis onset and cell degradation. RAB-35 binds CNT-1, an ARF-6 GTPase activating protein, and removes ARF-6, a degradation inhibitor, from phagosome membranes. This facilitates phosphatidylinositol-4,5-bisphosphate removal from phagosome membranes, promoting phagolysosome maturation. Our studies suggest that RAB-35 and ARF-6 drive a conserved program eliminating cells dying by LCD.

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Coscoy S, Baiz S, Octon J, Rhoné B, Perquis L, Tseng Q, Amblard F, Semetey V. (2018 Oct 16)

Microtopographies control the development of basal protrusions in epithelial sheets

Biointerphases : 13 : 041003 : DOI : 10.1116/1.5024601 Learn more
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Venzac B, Madoun R, Benarab T, Monnier S, Cayrac F, Myram S, Leconte L, Amblard F, Viovy JL, Descroix S, Coscoy S (2018 Oct 16)

Engineering small tubes with changes in diameter for the study of kidney cell organization

Biomicrofluidics : 12 : 024114 : DOI : 10.1063/1.5025027 Learn more
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