Cofilin recruitment and function during actin-mediated endocytosis dictated by actin nucleotide state.
Cofilin is the major mediator of actin filament turnover in vivo. However, the molecular mechanism of cofilin recruitment to actin networks during dynamic actin-mediated processes in living cells and cofilin's precise in vivo functions have not been determined. In this study, we analyzed the dynamics of fluorescently tagged cofilin and the role of cofilin-mediated actin turnover during endocytosis in Saccharomyces cerevisiae. In living cells, cofilin is not necessary for actin assembly on endocytic membranes but is recruited to molecularly aged adenosine diphosphate actin filaments and is necessary for their rapid disassembly. Defects in cofilin function alter the morphology of actin networks in vivo and reduce the rate of actin flux through actin networks. The consequences of decreasing actin flux are manifested by decreased but not blocked endocytic internalization at the plasma membrane and defects in late steps of membrane trafficking to the vacuole. These results suggest that cofilin-mediated actin filament flux is required for the multiple steps of endocytic trafficking.
Okreglak
Voytek
V
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.
Drubin
David G
DG
D008841:Actin Cytoskeleton; D051339:Actin Depolymerizing Factors / Q000378:metabolism*; D000199:Actins / Q000737:chemistry* / Q000378:metabolism*; D002268:Carboxypeptidases / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D004705:Endocytosis; D006867:Hydrolases / Q000378:metabolism; D008840:Microfilament Proteins / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation / Q000235:genetics; D009711:Nucleotides / Q000378:metabolism*; D011487:Protein Conformation; D021122:Protein Subunits / Q000378:metabolism; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D014617:Vacuoles / Q000201:enzymology
C062612:ABP1 protein, S cerevisiae; D051339:Actin Depolymerizing Factors; D000199:Actins; D008840:Microfilament Proteins; D009711:Nucleotides; D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; D006867:Hydrolases; D002268:Carboxypeptidases; C520242:CPY protein, S cerevisiae