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PMID: 9128251 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

High rates of actin filament turnover in budding yeast and roles for actin in establishment and maintenance of cell polarity revealed using the actin inhibitor latrunculin-A.

The Journal of cell biology ·Vol. 137 ·No. 2 ·1997-04-21 ·Pages 399-416

Ayscough KR, Stryker J, Pokala N, Sanders M, Crews P, Drubin DG

Abstract

We report that the actin assembly inhibitor latrunculin-A (LAT-A) causes complete disruption of the yeast actin cytoskeleton within 2-5 min, suggesting that although yeast are nonmotile, their actin filaments undergo rapid cycles of assembly and disassembly in vivo. Differences in the LAT-A sensitivities of strains carrying mutations in components of the actin cytoskeleton suggest that tropomyosin, fimbrin, capping protein, Sla2p, and Srv2p act to increase actin cytoskeleton stability, while End3p and Sla1p act to decrease stability. Identification of three LAT-A resistant actin mutants demonstrated that in vivo effects of LAT-A are due specifically to impairment of actin function and implicated a region on the three-dimensional actin structure as the LAT-A binding site. LAT-A was used to determine which of 19 different proteins implicated in cell polarity development require actin to achieve polarized localization. Results show that at least two molecular pathways, one actin-dependent and the other actin-independent, underlie polarity development. The actin-dependent pathway localizes secretory vesicles and a putative vesicle docking complex to sites of cell surface growth, providing an explanation for the dependence of polarized cell surface growth on actin function. Unexpectedly, several proteins that function with actin during cell polarity development, including an unconventional myosin (Myo2p), calmodulin, and an actin-interacting protein (Bud6/Aip3p), achieved polarized localization by an actin-independent pathway, revealing interdependence among cell polarity pathways. Finally, transient actin depolymerization caused many cells to abandon one bud site or mating projection and to initiate growth at a second site. Thus, actin filaments are also required for maintenance of an axis of cell polarity.

MeSH Terms
Actin Cytoskeleton/metabolism Actins/antagonists & inhibitors,metabolism Bridged Bicyclo Compounds, Heterocyclic/pharmacology Cell Cycle Proteins/analysis Cell Polarity/physiology Endocytosis/drug effects Fungal Proteins/analysis Resting Phase, Cell Cycle Saccharomyces cerevisiae/chemistry,cytology,drug effects Thiazoles/pharmacology Thiazolidines
Chemicals
Actins Bridged Bicyclo Compounds, Heterocyclic Cell Cycle Proteins Fungal Proteins Thiazoles Thiazolidines latrunculin A
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Ayscough K R
Department of Molecular and Cell Biology, University of California, Berkeley 94720-3202, USA.
Stryker J
Pokala N
Sanders M
Crews P
Drubin D G
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.137.2.399
Published
1997-04-21
Pages
399-416
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2139767
Subset
IM
Grants
NCI NIH HHS · CA47135 · United States
NIGMS NIH HHS · GM-42759 · United States
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1083/jcb.137.2.399 PMC Full Text View journal details
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