Microtubules under mechanical pressure can breach dense actin networks

The crosstalk between actin network and microtubules is key to the establishment of cell polarity. It ensures that the asymmetry of actin architec ture along cell periphery directs the organization of microtubules in cell interior. In particular, the way the two networks are physically inter-twined regulates the spatial organization and the distribution of forces in the microtubule network. While their biochemical crosstalk is getting uncovered, their mechanical crosstalk is still poorly understood. Here we designed an in vitro reconstitution assay to study the physical interaction between dynamic microtubules with various structures made of actin fil aments. We found that microtubules can align and move by their polymerization force along linear bundles of actin filaments. But they cannot enter dense and branched actin meshworks, such as those found in the lamellipodium along cell periphery. However, when microtubules are immobilized, by their crosslinking with actin structures or others means, the force of polymerization builds up pressure in the microtubules that is sufficient to allow them to breach and penetrate these dense actin meshworks. This mechanism may explain the final progression of microtubules up to cell periphery through the denser parts of the actin network.

[1]  A. Mogilner,et al.  Friction patterns guide actin network contraction , 2022, bioRxiv.

[2]  Alessandro Dema,et al.  Growth cone advance requires EB1 as revealed by genomic replacement with a light-sensitive variant , 2022, bioRxiv.

[3]  L. Blanchoin,et al.  Actin architecture steers microtubules in active cytoskeletal composite , 2022, bioRxiv.

[4]  L. Blanchoin,et al.  Actin network architecture can ensure robust centering or sensitive decentering of the centrosome , 2022, The EMBO journal.

[5]  A. Akhmanova,et al.  Mechanisms of microtubule organization in differentiated animal cells , 2022, Nature Reviews Molecular Cell Biology.

[6]  L. Blanchoin,et al.  Actin–microtubule dynamic composite forms responsive active matter with memory , 2022, bioRxiv.

[7]  K. Rottner,et al.  A barbed end interference mechanism reveals how capping protein promotes nucleation in branched actin networks , 2021, Nature Communications.

[8]  P. R. ten Wolde,et al.  Cross-linkers at growing microtubule ends generate forces that drive actin transport , 2021, bioRxiv.

[9]  S. Köster,et al.  Vimentin intermediate filaments stabilize dynamic microtubules by direct interactions , 2020, Nature Communications.

[10]  Michael J Rust,et al.  Actin and microtubule crosslinkers tune mobility and control co-localization in a composite cytoskeletal network. , 2020, Soft matter.

[11]  A. Mogilner,et al.  Stress fibers are embedded in a contractile cortical network , 2020, bioRxiv.

[12]  M. Bornens,et al.  Acto-myosin network geometry defines centrosome position , 2020, Current Biology.

[13]  C. Janke,et al.  Microtubule-Associated Proteins: Structuring the Cytoskeleton. , 2019, Trends in cell biology.

[14]  C. Hoogenraad,et al.  Cytolinker Gas2L1 regulates axon morphology through microtubule‐modulated actin stabilization , 2019, EMBO reports.

[15]  L. Blanchoin,et al.  Actin filaments regulate microtubule growth at the centrosome , 2019, The EMBO journal.

[16]  Christophe Leterrier,et al.  About samples, giving examples: Optimized Single Molecule Localization Microscopy , 2019, bioRxiv.

[17]  T. Mitchison Colloid osmotic parameterization and measurement of subcellular crowding , 2019, Molecular biology of the cell.

[18]  G. Koenderink,et al.  Actin–microtubule crosstalk in cell biology , 2018, Nature Reviews Molecular Cell Biology.

[19]  L. Blanchoin,et al.  Actin-Network Architecture Regulates Microtubule Dynamics , 2018, Current Biology.

[20]  E. Debold,et al.  Active Self-Organization of Actin-Microtubule Composite Self-Propelled Rods , 2018, Front. Phys..

[21]  E. Mandelkow,et al.  Multivalent cross-linking of actin filaments and microtubules through the microtubule-associated protein Tau , 2017, Nature Communications.

[22]  A. Mogilner,et al.  Network heterogeneity regulates steering in actin-based motility , 2017, Nature Communications.

[23]  X. Gidrol,et al.  Microtubule stabilization drives 3D centrosome migration to initiate primary ciliogenesis , 2017, bioRxiv.

[24]  G. Blin,et al.  Polarity Reversal by Centrosome Repositioning Primes Cell Scattering during Epithelial-to-Mesenchymal Transition. , 2017, Developmental cell.

[25]  L. Hodgson,et al.  Mesenchymal Cell Invasion Requires Cooperative Regulation of Persistent Microtubule Growth by SLAIN2 and CLASP1. , 2016, Developmental cell.

[26]  Yanan Yu,et al.  The CAMSAP3-ACF7 Complex Couples Noncentrosomal Microtubules with Actin Filaments to Coordinate Their Dynamics. , 2016, Developmental cell.

[27]  Clare M. Waterman,et al.  The FAK–Arp2/3 interaction promotes leading edge advance and haptosensing by coupling nascent adhesions to lamellipodia actin , 2016, Molecular biology of the cell.

[28]  V. Studer,et al.  Multiprotein Printing by Light‐Induced Molecular Adsorption , 2016, Advanced materials.

[29]  M. Valentine,et al.  The +TIP coordinating protein EB1 is highly dynamic and diffusive on microtubules, sensitive to GTP analog, ionic strength, and EB1 concentration , 2016, Cytoskeleton.

[30]  L. Blanchoin,et al.  Tau co-organizes dynamic microtubule and actin networks , 2015, Scientific Reports.

[31]  M. Steinmetz,et al.  Actin–microtubule coordination at growing microtubule ends , 2014, Nature Communications.

[32]  M. Théry,et al.  Quantification of MAP and molecular motor activities on geometrically controlled microtubule networks , 2013, Cytoskeleton.

[33]  L. Blanchoin,et al.  Nucleation geometry governs ordered actin networks structures. , 2010, Nature materials.

[34]  A. Malliri,et al.  The Rac activator STEF (Tiam2) regulates cell migration by microtubule‐mediated focal adhesion disassembly , 2010, EMBO reports.

[35]  M. Bornens Organelle positioning and cell polarity , 2008, Nature Reviews Molecular Cell Biology.

[36]  Elaine Fuchs,et al.  ACF7 Regulates Cytoskeletal-Focal Adhesion Dynamics and Migration and Has ATPase Activity , 2008, Cell.

[37]  G. Danuser,et al.  Coordination of actin filament and microtubule dynamics during neurite outgrowth. , 2008, Developmental cell.

[38]  Gaudenz Danuser,et al.  Filopodial actin bundles are not necessary for microtubule advance into the peripheral domain of Aplysia neuronal growth cones , 2007, Nature Cell Biology.

[39]  C. Waterman-Storer,et al.  Conserved microtubule–actin interactions in cell movement and morphogenesis , 2003, Nature Cell Biology.

[40]  Marileen Dogterom,et al.  Dynamic instability of microtubules is regulated by force , 2003, The Journal of cell biology.

[41]  Gary M. Bokoch,et al.  Regulation of leading edge microtubule and actin dynamics downstream of Rac1 , 2003, The Journal of cell biology.

[42]  George Oster,et al.  Force generation by actin polymerization II: the elastic ratchet and tethered filaments. , 2003, Biophysical journal.

[43]  Gary G. Borisy,et al.  Mechanism of filopodia initiation by reorganization of a dendritic network , 2003, The Journal of cell biology.

[44]  C. Waterman-Storer,et al.  Converging Populations of F-Actin Promote Breakage of Associated Microtubules to Spatially Regulate Microtubule Turnover in Migrating Cells , 2002, Current Biology.

[45]  C. Waterman-Storer,et al.  Dual-wavelength fluorescent speckle microscopy reveals coupling of microtubule and actin movements in migrating cells , 2002, The Journal of cell biology.

[46]  A. W. Schaefer,et al.  Filopodia and actin arcs guide the assembly and transport of two populations of microtubules with unique dynamic parameters in neuronal growth cones , 2002, The Journal of cell biology.

[47]  C. Cohan,et al.  Focal loss of actin bundles causes microtubule redistribution and growth cone turning , 2002, The Journal of cell biology.

[48]  G. Bokoch,et al.  Nucleotide exchange factor GEF-H1 mediates cross-talk between microtubules and the actin cytoskeleton , 2002, Nature Cell Biology.

[49]  B. Hinz,et al.  Actin-dependent lamellipodia formation and microtubule-dependent tail retraction control-directed cell migration. , 2000, Molecular biology of the cell.

[50]  P. Sansonetti,et al.  Activation of the Cdc42 Effector N-Wasp by the Shigella flexneri Icsa Protein Promotes Actin Nucleation by Arp2/3 Complex and Bacterial Actin-Based Motility , 1999, The Journal of cell biology.

[51]  E. Salmon,et al.  Actomyosin-based Retrograde Flow of Microtubules in the Lamella of Migrating Epithelial Cells Influences Microtubule Dynamic Instability and Turnover and Is Associated with Microtubule Breakage and Treadmilling , 1997, The Journal of cell biology.

[52]  A. Fellous,et al.  Characterization of a 100-kDa heat-stable microtubule-associated protein from higher plants. , 1994, European journal of biochemistry.

[53]  M. Shelanski CHEMISTRY OF THE FILAMENTS AND TUBULES OF BRAIN , 1973, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[54]  J. Spudich,et al.  The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. , 1971, The Journal of biological chemistry.

[55]  Laurent Blanchoin,et al.  Actin dynamics, architecture, and mechanics in cell motility. , 2014, Physiological reviews.

[56]  A. Hyman,et al.  Preparation of modified tubulins. , 1991, Methods in enzymology.