Cofilin promotes stimulus-induced lamellipodium formation by generating an abundant supply of actin monomers

Cofilin stimulates actin filament disassembly and accelerates actin filament turnover. Cofilin is also involved in stimulus-induced actin filament assembly during lamellipodium formation. However, it is not clear whether this occurs by replenishing the actin monomer pool, through filament disassembly, or by creating free barbed ends, through its severing activity. Using photoactivatable Dronpa-actin, we show that cofilin is involved in producing more than half of all cytoplasmic actin monomers and that the rate of actin monomer incorporation into the tip of the lamellipodium is dependent on the size of this actin monomer pool. Finally, in cofilin-depleted cells, stimulus-induced actin monomer incorporation at the cell periphery is attenuated, but the incorporation of microinjected actin monomers is not. We propose that cofilin contributes to stimulus-induced actin filament assembly and lamellipodium extension by supplying an abundant pool of cytoplasmic actin monomers.

[1]  Masahiro Yamamoto,et al.  Spatial and temporal regulation of cofilin activity by LIM kinase and Slingshot is critical for directional cell migration , 2005, The Journal of cell biology.

[2]  J. Condeelis,et al.  Cofilin takes the lead , 2005, Journal of Cell Science.

[3]  A. Miyawaki,et al.  Regulated Fast Nucleocytoplasmic Shuttling Observed by Reversible Protein Highlighting , 2004, Science.

[4]  P. Lappalainen,et al.  Actin-depolymerizing factor and cofilin-1 play overlapping roles in promoting rapid F-actin depolymerization in mammalian nonmuscle cells. , 2004, Molecular biology of the cell.

[5]  Céline Revenu,et al.  The co-workers of actin filaments: from cell structures to signals , 2004, Nature Reviews Molecular Cell Biology.

[6]  John S. Condeelis,et al.  Cofilin Promotes Actin Polymerization and Defines the Direction of Cell Motility , 2004, Science.

[7]  T. Uemura,et al.  Cell Cycle-associated Changes in Slingshot Phosphatase Activity and Roles in Cytokinesis in Animal Cells* , 2003, Journal of Biological Chemistry.

[8]  T. Uemura,et al.  Control of Growth Cone Motility and Morphology by LIM Kinase and Slingshot via Phosphorylation and Dephosphorylation of Cofilin , 2003, The Journal of Neuroscience.

[9]  Thomas D Pollard,et al.  Cellular Motility Driven by Assembly and Disassembly of Actin Filaments , 2003, Cell.

[10]  I. Yahara,et al.  The actin-severing activity of cofilin is exerted by the interplay of three distinct sites on cofilin and essential for cell viability. , 2002, The Biochemical journal.

[11]  T. Uemura,et al.  Control of Actin Reorganization by Slingshot, a Family of Phosphatases that Dephosphorylate ADF/Cofilin , 2002, Cell.

[12]  J. Condeelis,et al.  How is actin polymerization nucleated in vivo? , 2001, Trends in cell biology.

[13]  Marie-France Carlier,et al.  Mechanism of Actin-Based Motility , 2001, Science.

[14]  H. Miki,et al.  WASP and WAVE family proteins: key molecules for rapid rearrangement of cortical actin filaments and cell movement. , 2001, Journal of cell science.

[15]  K. Gunsalus,et al.  Cofilin/ADF is required for cell motility during Drosophila ovary development and oogenesis , 2001, Nature Cell Biology.

[16]  D. Lawrence,et al.  Phosphorylation of Adf/Cofilin Abolishes Egf-Induced Actin Nucleation at the Leading Edge and Subsequent Lamellipod Extension , 2000, The Journal of cell biology.

[17]  Amanda Y. Chan,et al.  Role of Cofilin in Epidermal Growth Factor–Stimulated Actin Polymerization and Lamellipod Protrusion , 2000, The Journal of cell biology.

[18]  I. Yahara,et al.  Two activities of cofilin, severing and accelerating directional depolymerization of actin filaments, are affected differentially by mutations around the actin‐binding helix , 1999, The EMBO journal.

[19]  L. Cramer Role of actin-filament disassembly in lamellipodium protrusion in motile cells revealed using the drug jasplakinolide , 1999, Current Biology.

[20]  J. Bamburg,et al.  Putting a new twist on actin: ADF/cofilins modulate actin dynamics. , 1999, Trends in cell biology.

[21]  J H Hartwig,et al.  Simultaneous measurements of actin filament turnover, filament fraction, and monomer diffusion in endothelial cells. , 1998, Biophysical journal.

[22]  E. Nishida,et al.  Cofilin phosphorylation by LIM-kinase 1 and its role in Rac-mediated actin reorganization , 1998, Nature.

[23]  P. Caroni,et al.  Regulation of actin dynamics through phosphorylation of cofilin by LIM-kinase , 1998, Nature.

[24]  S. Maciver How ADF/cofilin depolymerizes actin filaments. , 1998, Current opinion in cell biology.

[25]  J. Segall,et al.  EGF stimulates an increase in actin nucleation and filament number at the leading edge of the lamellipod in mammary adenocarcinoma cells. , 1998, Journal of cell science.

[26]  P. Lappalainen,et al.  Cofilin promotes rapid actin filament turnover in vivo , 1997, Nature.

[27]  Marie-France Carlier,et al.  Actin Depolymerizing Factor (ADF/Cofilin) Enhances the Rate of Filament Turnover: Implication in Actin-based Motility , 1997, The Journal of cell biology.

[28]  T. Mitchison,et al.  Xenopus Actin Depolymerizing Factor/Cofilin (XAC) Is Responsible for the Turnover of Actin Filaments in Listeria monocytogenes Tails , 1997, The Journal of cell biology.

[29]  K. Gunsalus,et al.  Mutations in twinstar, a Drosophila gene encoding a cofilin/ADF homologue, result in defects in centrosome migration and cytokinesis , 1995, The Journal of cell biology.

[30]  D. Drubin,et al.  The ADF/cofilin proteins: stimulus-responsive modulators of actin dynamics. , 1995, Molecular biology of the cell.

[31]  T. Mitchison,et al.  Comparison of actin and cell surface dynamics in motile fibroblasts , 1992, The Journal of cell biology.

[32]  Julie A. Theriot,et al.  Actin microfilament dynamics in locomoting cells , 1991, Nature.

[33]  T D Pollard,et al.  Rate constants for the reactions of ATP- and ADP-actin with the ends of actin filaments , 1986, The Journal of cell biology.

[34]  D. Taylor,et al.  Probing the mechanism of incorporation of fluorescently labeled actin into stress fibers , 1986, The Journal of cell biology.

[35]  T. Pollard,et al.  Cellular Motility Driven by Assembly and Disassembly of Actin Filaments , 2003, Cell.

[36]  T. Mitchison,et al.  Actin dynamics in vivo. , 1997, Current opinion in cell biology.