Yeast actin patches are networks of branched actin filaments.
Cortical actin patches are the most prominent actin structure in budding and fission yeast. Patches assemble, move, and disassemble rapidly. We investigated the mechanisms underlying patch actin assembly and motility by studying actin filament ultrastructure within a patch. Actin patches were partially purified from Saccharomyces cerevisiae and examined by negative-stain electron microscopy (EM). To identify patches in the EM, we correlated fluorescence and EM images of GFP-labeled patches. Patches contained a network of actin filaments with branches characteristic of Arp2/3 complex. An average patch contained 85 filaments. The average filament was only 50-nm (20 actin subunits) long, and the filament to branch ratio was 3:1. Patches lacking Sac6/fimbrin were unstable, and patches lacking capping protein were relatively normal. Our results are consistent with Arp2/3 complex-mediated actin polymerization driving yeast actin patch assembly and motility, as described by a variation of the dendritic nucleation model.
Young
Michael E
ME
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110, USA.
Cooper
John A
JA
Bridgman
Paul C
PC
D008841:Actin Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure*; D051377:Actin-Related Protein 2; D000199:Actins / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure*; D002478:Cells, Cultured; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins / Q000172:deficiency / Q000235:genetics; D008840:Microfilament Proteins / Q000172:deficiency / Q000235:genetics; D008854:Microscopy, Electron; D008954:Models, Biological; D009154:Mutation / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
C477632:ARP2 protein, S cerevisiae; D051377:Actin-Related Protein 2; D000199:Actins; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D029701:Saccharomyces cerevisiae Proteins; C026742:plastin; D049452:Green Fluorescent Proteins