arp23 branched actin network mediates filopodia-like bundles formation in vitroarp23支肌动蛋白网络协调filopodia-like包体外形成.pdfVIP

Arp2/3 Branched Actin Network Mediates Filopodia-Like Bundles Formation In Vitro 1 2 1 2 1 Yaron Ideses , Yifat Brill-Karniely , Lior Haviv , Avinoam Ben-Shaul , Anne Bernheim-Groswasser * 1 Ben-Gurion University of the Negev, Beer-Sheva, Israel, 2 The Hebrew University, Jerusalem, Israel Abstract During cellular migration, regulated actin assembly takes place at the cell leading edge, with continuous disassembly deeper in the cell interior. Actin polymerization at the plasma membrane results in the extension of cellular protrusions in the form of lamellipodia and filopodia. To understand how cells regulate the transformation of lamellipodia into filopodia, and to determine the major factors that control their transition, we studied actin self-assembly in the presence of Arp2/3 complex, WASp-VCA and fascin, the major proteins participating in the assembly of lamellipodia and filopodia. We show that in the early stages of actin polymerization fascin is passive while Arp2/3 mediates the formation of dense and highly branched aster-like networks of actin. Once filaments in the periphery of an aster get long enough, fascin becomes active, linking the filaments into bundles which emanate radially from the aster’s surface, resulting in the formation of star-like structures. We show that the number of bundles nucleated per star, as well as their thickness and length, is controlled by the initial concentration of Arp2/3 complex ([Arp2/3]). Specifically, we tested several values of [Arp2/3] and found that for given initial concentrations of actin and fascin, the number of bundles per star, as well as their length and thickness are larger when [Arp2/3] is lower. Our experimental findings can be interpreted and explained using a th