Yeast Genetics and Molecular Biology 2000
University of Washington
Seattle, Washington USA
July 2000


Name: Kemp, Hilary
Mailing Address: Institute of Molecular Biology, University of Oregon, 297 Klamath Hall, Eugene, OR 97403, USA
Email Address: hkemp@molbio.uoregon.edu
Phone & FAX numbers: 541-346-5158 &

#128"

Proteins that interact with Far3p are required for pheromone-mediated G1 arrest.
Hilary Kemp (1), Becky Drees (2), Stan Fields (2), George F. Sprague, Jr. (1)
(1) Institute of Molecular Biology, University of Oregon, 297 Klamath Hall, Eugene, OR 97403, USA; (2) Howard Hughes Medical Institute, University of Washington, Box 357360, Seattle, WA 98195-7360, USA

Activation of the yeast pheromone response pathway has three key effects: transcription induction of mating specific genes; G1 arrest; and shmoo formation. FAR3 is one of several genes required for G1 arrest. However Far3p does not appear to participate in any of the previously characterized mechanisms by which pheromone response gives rise to G1 arrest. In an effort to learn about the mechanism by which Far3p acts, we have carried out two-hybrid screens. We have identified nine putative Far3p interactors. Three of these were initially selected for further characterization. As is observed for far3 mutants, null mutants of FRG1, 2, or 3 (FAR3 interactor Required for G1 arrest) recover from pheromone-induced G1 arrest more rapidly than the wild type. Over-expression studies show that high levels of Far3p or Frg2p can restore G1 arrest in far3, frg1, frg2, or frg3 mutants. The effect of over-expressing Frg1p or Frg3p has not yet been examined. Conversely, over-expressing truncated versions of Frg1p or Frg3p causes a G1 arrest defect in the wild-type. Truncated versions of Far3p or Frg2p have not yet been tested. Subcellular fractionation of Far3p, Frg1p, and Frg2p reveals that all three proteins are present in the same fraction. Localization of Frg3p is underway. In toto, these data support the notion that Far3p and its interactors function together to regulate G1 arrest.


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