Nuclear export of Far1p in response to pheromones requires the export receptor Msn5p/Ste21p (original) (raw)

  1. Marc Blondel,
  2. Paula M. Alepuz,
  3. Linda S. Huang,
  4. Shai Shaham,
  5. Gustav Ammerer, and
  6. Matthias Peter
  7. Swiss Institute for Experimental Cancer Research (ISREC), 1066 Epalinges/VD, Switzerland; Vienna Biocenter, Institute of Biochemistry and Molecular Cell Biology, University of Vienna and Ludwig Boltzmann–Forschungsstelle für Biochemie, 1030 Vienna, Austria; Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, California 94143-0448 USA

Abstract

Far1p is a bifunctional protein that is required to arrest the cell cycle and to establish cell polarity during yeast mating. Far1p is localized predominantly in the nucleus but accumulates in the cytoplasm in cells exposed to pheromones. Here we show that Far1p functions in both subcellular compartments: nuclear Far1p is required to arrest the cell cycle, whereas cytoplasmic Far1p is involved in the establishment of cell polarity. The subcellular localization of Far1p is regulated by two mechanisms: (1) Far1p contains a functional bipartite nuclear localization signal (NLS), and (2) Far1p is exported from the nucleus by Msn5p/Ste21p, a member of the exportin family. Cells deleted for Msn5p/Ste21p failed to export Far1p in response to pheromones, whereas overexpression of Msn5p/Ste21p was sufficient to accumulate Far1p in the cytoplasm in the absence of pheromones. Msn5p/Ste21p was localized in the nucleus and interacted with Far1p in a manner dependent on GTP-bound Gsp1p. Two-hybrid analysis identified a small fragment within Far1p that is necessary and sufficient for binding to Msn5p/Ste21p, and is also required to export Far1p in vivo. Finally, similar to Δ_msn5/ste21_ strains, cells expressing a mutant Far1p, which can no longer be exported, exhibit a mating defect, but are able to arrest their cell cycle in response to pheromones. Taken together, our results suggest that nuclear export of Far1p by Msn5p/Ste21p coordinates the two separable functions of Far1p during mating.

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