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A more recent version of this article appeared on March 1, 2003
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Submitted on May 30, 2002
Revised on October 1, 2002
Accepted on October 8, 2002
1 Department of Cell Biology and Oncology, Istituto di Ricerche Farmacologiche Mario Negri, Consorzio Mario Negri Sud, S. Maria Imbaro (Chieti), Italy
2 Department of Cell Biology and Oncology, Istituto di Ricerche Farmacologiche Mario Negri, Consorzio Mario Negri Sud, S. Maria Imbaro (Chieti), Italy; and Endocrine Unit, Department of Internal Medicine, Ospedale Civile "Renzetti", Lanciano (Chieti), Italy
3 Department of Biochemistry and Molecular Biology, Center for Basic Research in Digestive Diseases, Mayo Clinic, Rochester MN, USA
* Corresponding author. E-mail address: luini{at}dcbo.negrisud.it.
* Corresponding author. E-mail address: buccione{at}dcbo.negrisud.it.
The degradation of extracellular matrix (ECM) by matrix metalloproteases (MMPs) is crucial in physiological and pathological cell invasion alike. Degradation occurs at specific sites where invasive cells make contact with the ECM via specialized plasma membrane protrusions termed invadopodia. Here we show that the dynamin 2, a GTPase implicated in the control of actin-driven cytoskeletal remodeling events and membrane transport is necessary for focalized matrix degradation at invadopodia. Dynamin was inhibited by employing two approaches: 1) expression of dominant negative GTPase-impaired or proline-rich domain-deleted dynamin 2 mutants and 2) inhibition of the dynamin regulator calcineurin by Cyclosporin A. In both cases, the number and extension of ECM degradation foci were drastically reduced. To understand the site and mechanism of dynamin action, the cellular structures devoted to ECM degradation were analyzed by correlative confocal light-electron microscopy. Invadopodia were found to be organized into a previously undescribed ECM-degradation structure (EDS) consisting of a large invagination of the ventral plasma membrane surface in close spatial relationship with the Golgi complex. Dynamin 2 appeared to be concentrated at invadopodia.
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