coordination of membrane and actin cytoskeleton dynamics during filopodia protrusion协调膜和肌动蛋白细胞骨架动力学在丝状伪足突出.pdfVIP

Coordination of Membrane and Actin Cytoskeleton Dynamics during Filopodia Protrusion 1 1 2 2 1 Changsong Yang , Matthew Hoelzle , Andrea Disanza , Giorgio Scita , Tatyana Svitkina * 1 Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America, 2 The Italian Foundation for Cancer Research (FIRC), Institute for Molecular Oncology and Department of Experimental Oncology, European Institute of Oncology, Milan, Italy Abstract Leading edge protrusion of migrating cells involves tightly coordinated changes in the plasma membrane and actin cytoskeleton. It remains unclear whether polymerizing actin filaments push and deform the membrane, or membrane deformation occurs independently and is subsequently stabilized by actin filaments. To address this question, we employed an ability of the membrane-binding I-BAR domain of IRSp53 to uncouple the membrane and actin dynamics and to induce filopodia in expressing cells. Using time-lapse imaging and electron microscopy of IRSp53-I-BAR-expressing B16F1 melanoma cells, we demonstrate that cells are not able to protrude or maintain durable long extensions without actin filaments in their interior, but I-BAR-dependent membrane deformation can create a small and transient space at filopodial tips that is subsequently filled with actin filaments. Moreover, the expressed I-BAR domain forms a submembranous coat that may structurally support these transient actin-free protrusions until they are further stabilized by the actin cytoskeleton. Actin filaments in the I-BAR-induced filopodia, in contrast to normal filopodia, do not have a uniform length, are less abundant, poorly bundled, an