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Publications de l’équipe Dynamique de la membrane et du cytosquelette Année de publication : 2014 Guillaume Montagnac, Philippe Chavrier (2014 Feb 28) [When microtubules meet clathrin-coated pits]. Mé decine sciences : M/S : 130-3 : DOI : 10.1051/medsci/20143002005 Résumé Année de publication : 2013 Pedro Monteiro, Carine Rossé, Antonio Castro-Castro, Marie Irondelle, Emilie Lagoutte, Perrine Paul-Gilloteaux, Claire Desnos, Etienne Formstecher, François Darchen, David Perrais, Alexis Gautreau, Maud Hertzog, Philippe Chavrier (2013 Dec 18) Endosomal WASH and exocyst complexes control exocytosis of MT1-MMP at invadopodia. The Journal of cell biology : 1063-79 Résumé Remodeling of the extracellular matrix by carcinoma cells during metastatic dissemination requires formation of actin-based protrusions of the plasma membrane called invadopodia, where the trans-membrane type 1 matrix metalloproteinase (MT1-MMP) accumulates. Here, we describe an interaction between the exocyst complex and the endosomal Arp2/3 activator Wiskott-Aldrich syndrome protein and Scar homolog (WASH) on MT1MMP–containing late endosomes in invasive breast carcinoma cells. We found that WASH and exocyst are required for matrix degradation by an exocytic mechanism that involves tubular connections between MT1-MMP–positive late endosomes and the plasma membrane in contact with the matrix. This ensures focal delivery of MT1-MMP and supports pericellular matrix degradation and tumor cell invasion into different pathologically relevant matrix environments. Our data suggest a general mechanism used by tumor cells to breach the basement membrane and for invasive migration through fibrous collagen-enriched tissues surrounding the tumor. Maud Hertzog, Pedro Monteiro, Gaëlle Le Dez, Philippe Chavrier (2013 Jan 10) Exo70 subunit of the exocyst complex is involved in adhesion-dependent trafficking of caveolin-1. PloS one : e52627 : DOI : 10.1371/journal.pone.0052627 Résumé Caveolae are specialized domains of the plasma membrane, which play key roles in signaling, endocytosis and mechanosensing. Using total internal reflection fluorescent microscopy (TIRF-M), we observe that the exocyst subunit Exo70 forms punctuate structures INSTITUT CURIE, 20 rue d’Ulm, 75248 Paris Cedex 05, France | 1 Publications de l’équipe Dynamique de la membrane et du cytosquelette at the plasma membrane and partially localizes with caveolin-1, the main component of caveolae. Upon cell detachment, we found that Exo70 accumulates with caveolin-1-positive vesicular structures. Upon cell re-adhesion, caveolin-1 traffics back to the plasma membrane in a multistep process involving microtubules and actin cytoskeleton. In addition, silencing of Exo70 redirects caveolin-1 to focal adhesions identified by markers such as α5 integrin or vinculin. Based on these findings, we conclude that Exo70 is involved in caveolin-1 recycling to the plasma membrane during re-adhesion of the cells to the substratum. Année de publication : 2012 Antonio Castro-Castro, Carsten Janke, Guillaume Montagnac, Perrine Paul-Gilloteaux, Philippe Chavrier (2012 Dec 9) ATAT1/MEC-17 acetyltransferase and HDAC6 deacetylase control a balance of acetylation of alpha-tubulin and cortactin and regulate MT1-MMP trafficking and breast tumor cell invasion. European journal of cell biology : 950-60 : DOI : 10.1016/j.ejcb.2012.07.001 Résumé Invasive tumor cells use proteases to degrade and migrate through the stromal environment consisting of a 3D network of extracellular matrix macromolecules. In particular, MT1-MMP, a membrane-anchored metalloproteinase, is critical during cancer cell invasion. MT1-MMP is stored in endosomal compartments and then delivered to invadopodia, the specialized plasma membrane domains of invasive cancer cells endowed with extracellular matrixdegradation capacity. In macrophages, traffic of MT1-MMP vesicles to invadopodia-related podosomes requires microtubules. We previously found that in breast tumor MDA-MB-231 cells an increase of microtubule and cortactin acetylation upon inhibition of HDAC6 correlates with a decrease of matrix degradation and invasion in three-dimensional collagen I gel. Here, we investigated the role of the recently identified α-tubulin N-acetyltransferase 1 ATAT1 in invasive MDA-MB-231 cells. We found that the dynamics and distribution of MT1MMP-positive endosomes require regulation of acetylation levels. We observed that ATAT1 tubulin acetyltransferase binds and regulates cortactin acetylation levels. In addition, ATAT1 colocalizes with cortactin at the adherent surface of the cells and it is required for 2D migration and invasive migration of MDA-MB-231 cells in collagen matrix. All together, our data indicate that a balance of acetylation and deaceylation by ATAT1/HDAC6 enzymes with opposite activities regulates the migratory and invasive capacities of breast tumor cells. INSTITUT CURIE, 20 rue d’Ulm, 75248 Paris Cedex 05, France | 2