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First published online 15 March 2005
doi: 10.1242/jcs.02285

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WASP-related proteins, Abi1 and Ena/VASP are required for Listeria invasion induced by the Met receptor

¹ Unité des Interactions Bactéries-Cellules, Institut Pasteur, INSERM U604, INRA USC2020, 28 rue du Docteur Roux, 75724 Paris Cedex 15, France
² Division of Cancer Genomics, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, MInato-ku, Tokyo 108-8639, Japan, and PRESTO, Japan Science and Technology Agency (JST)
³ The European Institute of Oncology (IEO) and the FIRC Institute for Molecular Oncology (IFOM), Via Adamello 16, 20139 Milan, Italy
⁴ Department of Biology, Massachusetts Institute of Technology, 31 Ames Street, Cambridge, MA 02139-4307, USA
⁵ Department of Biochemistry, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan

^* Author for correspondence (e-mail: pcossart{at}pasteur.fr)

Accepted 25 January 2005

Internalisation of the pathogenic bacterium Listeria monocytogenes involves interactions between the invasion protein InlB and the hepatocyte growth factor receptor, Met. Using colocalisation studies, dominant-negative constructs and small interfering RNA (siRNA), we demonstrate a cell-type-dependent requirement for various WASP-related proteins in Listeria entry and InlB-induced membrane ruffling. The WAVE2 isoform is essential for InlB-induced cytoskeletal rearrangements in Vero cells. In HeLa cells, WAVE1, WAVE2 and N-WASP cooperate to promote these processes. Abi1, a key component of WAVE complexes, is recruited at the entry site in both cell types and its inactivation by RNA interference impairs InlB-mediated processes. Ena/VASP proteins also play a role in Listeria internalization, and their deregulation by sequestration or overexpression, modifies actin cups beneath entering particles. Taken together, these results identify the WAVE complex, N-WASP and Ena/VASP as key effectors of the Met signalling pathway and of Listeria entry and highlight the existence of redundant and/or cooperative functions among WASP-family members.

Key words: Listeria monocytogenes, Cytoskeleton, HGF, Actin, Rac, Cdc42, Phagocytosis

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