The Yeast V159N Actin Mutant Reveals Roles for Actin Dynamics In Vivo

doi:10.1083/jcb.142.5.1289

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© The Rockefeller University Press, 0021-9525/1998//1289 $5.00
The Journal of Cell Biology, Volume 142, Number 5, , 1998 1289-1299

Articles

The Yeast V159N Actin Mutant Reveals Roles for Actin Dynamics In Vivo

Lisa D. Belmont and David G. Drubin

Department of Molecular and Cell Biology, University of California, Berkeley, California 94720-3202

Actin with a Val 159 to Asn mutation (V159N) forms actin filamentsthat depolymerize slowly because of a failure to undergo aconformational change after inorganic phosphate release. Herewe demonstrate that expression of this actin results in reducedactin dynamics in vivo, and we make use of this property tostudy the roles of rapid actin filament turnover. Yeast strainsexpressing the V159N mutant (act1-159) as their only sourceof actin have larger cortical actin patches and more actin cablesthan wild-type yeast. Rapid actin dynamics are not essentialfor cortical actin patch motility or establishment of cellpolarity. However, fluid phase endocytosis is defective in act1-159 strains. act1-159 is synthetically lethal with cofilin and profilin mutants, supporting the conclusion that mutationsin all of these genes impair the polymerization/ depolymerizationcycle. In contrast, act1-159 partially suppresses the temperaturesensitivity of a tropomyosin mutant, and the loss of cytoplasmiccables seen in fimbrin, Mdm20p, and tropomyosin null mutants,suggesting filament stabilizing functions for these actin-binding proteins. Analysis of the cables in these double-mutant cellssupports a role for fimbrin in organizing cytoplasmic cablesand for Mdm20p and tropomyosin in excluding cofilin from thecables.

Key Words: actin • dynamics • ATP hydrolysis • yeast • cytoskeleton

Abbreviations used in this paper: F-actin, filamentous actin; LAT-A, latrunculin-A.

Address all correspondence to David G. Drubin, Department ofMolecular and Cell Biology, 401 Barker Hall, University of California,Berkeley, CA 94720-3202. Tel.: (510) 642-3692. Fax: (510) 642-6420.E-mail: drubin{at}uclink4.berkeley.edu

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