UMR168 – Physico-Chimie Curie Lab

Team Publications

Year of publication 2020

Pernier Julien, Morchain Antoine, Caorsi Valentina, Bertin Aurélie, Bousquet Hugo, Bassereau Patricia, Coudrier Evelyne (2020 Sep 7)

Myosin 1b Flattens and Prunes Branched Actin Filaments.

Journal of Cell Science : DOI : 10.1242/jcs.247403 Learn more
Summary

Abstract
Motile and morphological cellular processes require a spatially and temporally
coordinated branched actin network that is controlled by the activity of various regulatory
proteins including the Arp2/3 complex, profilin, cofilin and tropomyosin. We have previously
reported that myosin 1b regulates the density of the actin network in the growth cone. Using
in vitro F-actin gliding assays and total internal reflection fluorescence (TIRF) microscopy we
show in this report that this molecular motor flattens the Arp2/3-dependent actin branches
up to breaking them and reduces the probability to form new branches. This experiment
reveals that myosin 1b can produce force sufficient enough to break up the Arp2/3-mediated
actin junction. Together with the former in vivo studies, this work emphasizes the essential
role played by myosins in the architecture and in the dynamics of actin networks in different
cellular regions.

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Stec Natalia, Doerfel Katja, Hills-Muckey Kelly , Ettorre Victoria, Ercan Sevinc, Keil Wolfgang, Hammell Christopher (2020 Sep 3)

An Epigenetic Priming Mechanism Mediated by Nutrient Sensing Regulates Transcriptional Output

bioRxivAn Epigenetic Priming Mechanism Mediated by Nutrient Sensing Regulates Transcriptional Output : DOI : https://doi.org/10.1101/2020.09.01.278127 Learn more
Summary

While precise tuning of gene expression levels is critical for most developmental pathways, the mechanisms by which the transcriptional output of dosage-sensitive molecules is established or modulated by the environment remain poorly understood. Here, we provide a mechanistic framework for how the conserved transcription factor BLMP-1/Blimp1 operates as a pioneer factor to decompact chromatin near its target loci hours before transcriptional activation and by doing so, regulates both the duration and amplitude of subsequent target gene transcription. This priming mechanism is genetically separable from the mechanisms that establish the timing of transcriptional induction and functions to canalize aspects of cell-fate specification, animal size regulation, and molting. A key feature of the BLMP-1-dependent transcriptional priming mechanism is that chromatin decompaction is initially established during embryogenesis and maintained throughout larval development by nutrient sensing. This anticipatory mechanism integrates transcriptional output with environmental conditions and is essential for resuming normal temporal patterning after animals exit nutrient-mediated developmental arrests.

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Taveneau Cyntia, Blanc Rémi, Péhau-Arnaudet Gérard, Di Cicco Aurélie, Bertin Aurélie (2020 Jul 29)

Synergistic role of nucleotides and lipids for the self-assembly of Shs1 septin oligomers

Biochemical Journal : 477 : 2697-2714 : DOI : 10.1042/BCJ20200199 Learn more
Summary

Budding yeast septins are essential for cell division and polarity. Septins assemble as
palindromic linear octameric complexes. The function and ultra-structural organization of
septins are finely governed by their molecular polymorphism. In particular, in budding
yeast, the end subunit can stand either as Shs1 or Cdc11. We have dissected, here, for
the first time, the behavior of the Shs1 protomer bound to membranes at nanometer
resolution, in complex with the other septins. Using electron microscopy, we have shown
that on membranes, Shs1 protomers self-assemble into rings, bundles, filaments or twodimensional
gauzes. Using a set of specific mutants we have demonstrated a synergistic
role of both nucleotides and lipids for the organization and oligomerization of budding
yeast septins. Besides, cryo-electron tomography assays show that vesicles are
deformed by the interaction between Shs1 oligomers and lipids. The Shs1–Shs1 interface
is stabilized by the presence of phosphoinositides, allowing the visualization of micrometric
long filaments formed by Shs1 protomers. In addition, molecular modeling experiments
have revealed a potential molecular mechanism regarding the selectivity of septin
subunits for phosphoinositide lipids.

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Davidson PM, Battistella A, Déjardin T, Betz T, Plastino J, Cadot B, Borghi N, Sykes C (2020 Jul 3)

Nesprin-2 accumulates at the front of the nucleus during confined cell migration

EMBO Reports : DOI : 10.15252/embr.201949910 Learn more
Summary

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Cao Luyan, Yonis Amina, Vaghela Malti, Barriga Elias, Chugh Priyamvada, Smith Matthew, Maufront Julien, Lavoie Geneviève, Méant Antoine, Ferber Emma, Bovellan Miia, Alberts Art, Bertin Aurélie, Mayor Roberto, Paluch Eva, Roux Philippe, Jégou Antoine, Romet-Lemonne Guillaume, Charras Guillaume (2020 Jun 22)

SPIN90 associates with mDia1 and the Arp2/3 complex to regulate cortical actin organization

Nature Cell BiologyNature Cell Biology : DOI : 10.1038/s41556-020-0531-y Learn more
Summary

Cell shape is controlled by the submembranous cortex, an actomyosin network mainly generated by two actin nucleators: the Arp2/3 complex and the formin mDia1. Changes in relative nucleator activity may alter cortical organization, mechanics and cell shape. Here we investigate how nucleation-promoting factors mediate interactions between nucleators. In vitro, the nucleation-promoting factor SPIN90 promotes formation of unbranched filaments by Arp2/3, a process thought to provide the initial filament for generation of dendritic networks. Paradoxically, in cells, SPIN90 appears to favour a formin-dominated cortex. Our in vitro experiments reveal that this feature stems mainly from two mechanisms: efficient recruitment of mDia1 to SPIN90–Arp2/3 nucleated filaments and formation of a ternary SPIN90–Arp2/3–mDia1 complex that greatly enhances filament nucleation. Both mechanisms yield rapidly elongating filaments with mDia1 at their barbed ends and SPIN90–Arp2/3 at their pointed ends. Thus, in networks, SPIN90 lowers branching densities and increases the proportion of long filaments elongated by mDia1.

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Aurélie Bertin , Nicola de Franceschi , Eugenio de la Mora , Sourav Maiti, Maryam Alqabandi, Nolwen Miguet, Aurélie di Cicco, Wouter H. Roos, Stéphanie Mangenot , Winfried Weissenhorn, Patricia Bassereau (2020 May 29)

Human ESCRT-III polymers assemble on positively curved membranes and induce helical membrane tube formation

Nature Communications : 11 : 2663 : DOI : 10.1038/s41467-020-16368-5 Learn more
Summary

Endosomal sorting complexes for transport-III (ESCRT-III) assemble in vivo onto membranes with negative Gaussian curvature. How membrane shape influences ESCRT-III polymerization and how ESCRT-III shapes membranes is yet unclear. Human core ESCRT-III proteins, CHMP4B, CHMP2A, CHMP2B and CHMP3 are used to address this issue in vitro by combining membrane nanotube pulling experiments, cryo-electron tomography and AFM. We show that CHMP4B filaments preferentially bind to flat membranes or to tubes with positive mean curvature. Both CHMP2B and CHMP2A/CHMP3 assemble on positively curved membrane tubes. Combinations of CHMP4B/CHMP2B and CHMP4B/CHMP2A/CHMP3 are recruited to the neck of pulled membrane tubes and reshape vesicles into helical “corkscrewlike” membrane tubes. Sub-tomogram averaging reveals that the ESCRT-III filaments assemble parallel and locally perpendicular to the tube axis, highlighting the mechanical stresses imposed by ESCRT-III. Our results underline the versatile membrane remodeling activity of ESCRT-III that may be a general feature required for cellular membrane remodeling processes.

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Samandar Eweis D, Plastino J (2020 May 21)

Roles of actin in the morphogenesis of the early Caenorhabditis elegans embryo

International Journal of Molecular Sciences : DOI : 10.3390/ijms21103652 Learn more
Summary

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Gat S, Simon C, Campillo C, Bernheim-Groswasser A, Sykes C (2020 May 21)

Finger-like membrane protrusions are favored by heterogeneities in the actin network

Soft Matter : DOI : 10.1039/c9sm02444a Learn more
Summary

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Allard A, Bouzid M, Betz T, Simon C, Abou-Ghali M, Lemière J, Valentino F, Manzi J, Brochard-Wyart F, Guevorkian K, Plastino J, Lenz M, Campillo C*, Sykes C* (2020 Apr 22)

Actin modulates shape and mechanics of tubular membranes

Science Advances : DOI : 10.1126/sciadv.aaz3050 Learn more
Summary

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Year of publication 2019

Mijo Simunovic, Emma Evergren, Andrew Callan-Jones*, Patricia Bassereau* (2019 Oct 7)

Curving Cells Inside and Out: Roles of BAR Domain Proteins in Membrane Shaping and Its Cellular Implications.

Annual Review of Cell and Developmental Biology : 35 : DOI : 10.1146/annurev-cellbio-100617-060558 Learn more
Summary

Many cellular processes rely on precise and timely deformation of the cell membrane. While many proteins participate in membrane reshaping and scission, usually in highly specialized ways, Bin/amphiphysin/Rvs (BAR) domain proteins play a pervasive role, as they not only participate in many aspects of cell trafficking but also are highly versatile membrane remodelers. Subtle changes in the shape and size of the BAR domain can greatly impact the way in which BAR domain proteins interact with the membrane. Furthermore, the activity of BAR domain proteins can be tuned by external physical parameters, and so they behave differently depending on protein surface density, membrane tension, or membrane shape. These proteins can form 3D structures that mold the membrane and alter its liquid properties, even promoting scission under various circumstances. As such, BAR domain proteins have numerous roles within the cell. Endocytosis is among the most highly studied processes in which BAR domain proteins take on important roles. Over the years, a more complete picture has emerged in which BAR domain proteins are tied to almost all intracellular compartments; examples include endosomal sorting and tubular networks in the endoplasmic reticulum and T-tubules. These proteins also have a role in autophagy, and their activity has been linked with cancer. Here, we briefly review the history of BAR domain protein discovery, discuss the mechanisms by which BAR domain proteins induce curvature, and attempt to settle important controversies in the field. Finally, we review BAR domain proteins in the context of a cell, highlighting their emerging roles in cell signaling and organelle shaping.

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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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Moitrier Sarah, Pricoupenko Nastassia, Kerjouan Adèle, Oddou Christiane, Destaing Olivier, Battistella Aude, Silberzan Pascal, Bonnet Isabelle (2019 Sep 3)

Local light-activation of the Src oncoprotein in an epithelial monolayer promotes collective extrusion

Communications Physics : 2 : 98 : DOI : 10.1038/s42005-019-0198-5 Learn more
Summary

Transformed isolated cells are usually extruded from normal epithelia and subsequently eliminated. However, multicellular tumors outcompete healthy cells, highlighting the importance of collective effects. Here, we investigate this situation in vitro by controlling in space and time the activity of the Src oncoprotein within a normal Madin–Darby Canine Kidney (MDCK) epithelial cell monolayer. Using an optogenetics approach with cells expressing a synthetic light-sensitive version of Src (optoSrc), we reversibly trigger the oncogenic activity by exposing monolayers to well-defined light patterns. We show that small populations of activated optoSrc cells embedded in the non-transformed monolayer collectively extrude as a tridimensional aggregate and remain alive, while the surrounding normal cells migrate towards the exposed area. This phenomenon requires an interface between normal and transformed cells and is partially reversible. Traction forces show that Src- activated cells either actively extrude or are pushed out by the surrounding cells in a non- autonomous way.

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Zack Jarin, Feng-Ching Tsai, Aram Davtyan, Alexander J.Pak, Patricia Bassereau, Gregory A.Voth (2019 Aug 6)

Unusual Organization of I-BAR Proteins on Tubular and Vesicular Membranes.

Biophysical Journal : 117 : 553-562 : DOI : 10.1016/j.bpj.2019.06.025 Learn more
Summary

Protein-mediated membrane remodeling is a ubiquitous and critical process for proper cellular function. Inverse Bin/Amphiphysin/Rvs (I-BAR) domains drive local membrane deformation as a precursor to large-scale membrane remodeling. We employ a multiscale approach to provide the molecular mechanism of unusual I-BAR domain-driven membrane remodeling at a low protein surface concentration with near-atomistic detail. We generate a bottom-up coarse-grained model that demonstrates similar membrane-bound I-BAR domain aggregation behavior as our recent Mesoscopic Membrane with Explicit Proteins model. Together, these models bridge several length scales and reveal an aggregation behavior of I-BAR domains. We find that at low surface coverage (i.e., low bound protein density), I-BAR domains form transient, tip-to-tip strings on periodic flat membrane sheets. Inside of lipid bilayer tubules, we find linear aggregates parallel to the axis of the tubule. Finally, we find that I-BAR domains form tip-to-tip aggregates around the edges of membrane domes. These results are supported by in vitro experiments showing low curvature bulges surrounded by I-BAR domains on giant unilamellar vesicles. Overall, our models reveal new I-BAR domain aggregation behavior in membrane tubules and on the surface of vesicles at low surface concentration that add insight into how I-BAR domain proteins may contribute to certain aspects of membrane remodeling in cells.

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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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Nicola de Franceschi, Maryam Alqabandi, Winfried Weissenhorn, Patricia Bassereau (2019 Jul 5)

Dynamic and Sequential Protein Reconstitution on Negatively Curved Membranes by Giant Vesicles Fusion.

Bio-Protocol : 9 : e3294 : DOI : 10.21769/BioProtoc.3294 Learn more
Summary

In vitro investigation of the interaction between proteins and positively curved membranes can be performed using a classic nanotube pulling method. However, characterizing protein interaction with negatively curved membranes still represents a formidable challenge. Here, we describe our recently developed approach based on laser-triggered Giant Unilamellar Vesicles (GUVs) fusion. Our protocol allows sequential addition of proteins to a negatively curved membrane, while at the same time controlling the buffer composition, lipid composition and membrane tension. Moreover, this method does not require a step of protein detachment, greatly simplifying the process of protein encapsulation over existing methods.

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