The F-actin side binding activity of the Arp2/3 complex is essential for actin nucleation and lamellipod extension

Bailly, M., Ichetovkin, I., Grant, W., Zebda, N., Machesky, L. M. , Segall, J. E. and Condeelis, J. (2001) The F-actin side binding activity of the Arp2/3 complex is essential for actin nucleation and lamellipod extension. Current Biology, 11(8), pp. 620-625. (doi: 10.1016/S0960-9822(01)00152-X) (PMID:11369208)

Abstract

Most eukaryotic cells rely on localized actin polymerization to generate and sustain the protrusion activity necessary for cell movement 1, 2. Such protrusions are often in the form of a flat lamellipod with a leading edge composed of a dense network of actin filaments 3, 4. The Arp2/3 complex localizes within that network in vivo 3, 4 and nucleates actin polymerization and generates a branched network of actin filaments in vitro 5, 6, 7. The complex has thus been proposed to generate the actin network at the leading edge of crawling cells in vivo 3, 4, 8. However, the relative contributions of nucleation and branching to protrusive force are still unknown. We prepared antibodies to the p34 subunit of the Arp2/3 complex that selectively inhibit side binding of the complex to F-actin. We demonstrate that side binding is required for efficient nucleation and branching by the Arp2/3 complex in vitro. However, microinjection of these antibodies into cells specifically inhibits lamellipod extension without affecting the EGF-stimulated appearance of free barbed ends in situ. These results indicate that while the side binding activity of the Arp2/3 complex is required for nucleation in vitro and for protrusive force in vivo, it is not required for EGF-stimulated increases in free barbed ends in vivo. This suggests that the branching activity of the Arp2/3 complex is essential for lamellipod extension, while the generation of nucleation sites for actin polymerization is not sufficient.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Machesky, Professor Laura
• Authors: Bailly, M., Ichetovkin, I., Grant, W., Zebda, N., Machesky, L. M., Segall, J. E., and Condeelis, J.
• College/School: College of Medical Veterinary and Life Sciences > School of Cancer Sciences
• Journal Name: Current Biology
• Publisher: Cell Press
• ISSN: 0960-9822
• ISSN (Online): 1879-0445
• Published Online: 07 May 2001