A Synthetic Lethal Screen Identifies a Role for the Cortical Actin Patch/Endocytosis Complex in the Response to Nutrient Deprivation in Saccharomyces cerevisiae
Alison Care, Katherine A. Vousden, Katie M. Binley, Pippa Radcliffe, Janet Trevethick, Ilaria Mannazzu, Peter E. Sudbery

Abstract

Saccharomyces cerevisiae whi2Δ cells are unable to halt cell division in response to nutrient limitation and are sensitive to a wide variety of stresses. A synthetic lethal screen resulted in the isolation of siw mutants that had a phenotype similar to that of whi2Δ. Among these were mutations affecting SIW14, FEN2, SLT2, and THR4. Fluid-phase endocytosis is severely reduced or abolished in whi2Δ, siw14Δ, fen2Δ, and thr4Δ mutants. Furthermore, whi2Δ and siw14Δ mutants produce large actin clumps in stationary phase similar to those seen in prk1Δ ark1Δ mutants defective in protein kinases that regulate the actin cytoskeleton. Overexpression of SIW14 in a prk1Δ strain resulted in a loss of cortical actin patches and cables and was lethal. Overexpression of SIW14 also rescued the caffeine sensitivity of the slt2 mutant isolated in the screen, but this was not due to alteration of the phosphorylation state of Slt2. These observations suggest that endocytosis and the organization of the actin cytoskeleton are required for the proper response to nutrient limitation. This hypothesis is supported by the observation that rvs161Δ, sla1Δ, sla2Δ, vrp1Δ, ypt51Δ, ypt52Δ, and end3Δ mutations, which disrupt the organization of the actin cytoskeleton and/or reduce endocytosis, have a phenotype similar to that of whi2Δ mutants.

  • Received July 28, 2003.
  • Accepted October 27, 2003.
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