Actin filaments in yeast are unstable in the absence of capping protein or fimbrin.
Many actin-binding proteins affect filament assembly in vitro and localize with actin in vivo, but how their molecular actions contribute to filament assembly in vivo is not understood well. We report here that capping protein (CP) and fimbrin are both important for actin filament assembly in vivo in Saccharomyces cerevisiae, based on finding decreased actin filament assembly in CP and fimbrin mutants. We have also identified mutations in actin that enhance the CP phenotype and find that those mutants also have decreased actin filament assembly in vivo. In vitro, actin purified from some of these mutants is defective in polymerization or binding fimbrin. These findings support the conclusion that CP acts to stabilize actin filaments in vivo. This conclusion is particularly remarkable because it is the opposite of the conclusion drawn from recent studies in Dictyostelium (Hug, C., P.Y. Jay, I. Reddy, J.G. McNally, P.C. Bridgman, E.L. Elson, and J.A. Cooper. 1995. Cell. 81:591-600). In addition, we find that the unpolymerized pool of actin in yeast is very small relative to that found in higher cells, which suggests that actin filament assembly is less dynamic in yeast than higher cells.
Department of Cell Biology and Physiology, Washington University Medical School, St. Louis, Missouri, USA.
T S
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Karpova
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Tatchell
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JA
Cooper
D000199:Actins / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D000483:Alleles; D000818:Animals; D002352:Carrier Proteins / Q000502:physiology*; D003598:Cytoskeletal Proteins; D004023:Dictyostelium / Q000502:physiology; D005656:Fungal Proteins / Q000502:physiology; D008562:Membrane Glycoproteins / Q000502:physiology*; D008565:Membrane Proteins; D008840:Microfilament Proteins; D009154:Mutation / Q000502:physiology; D010957:Plasmids; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins
D000199:Actins; C090135:Cap2 protein, S cerevisiae; D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; D005656:Fungal Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D008840:Microfilament Proteins; D029701:Saccharomyces cerevisiae Proteins; C026742:plastin