title: Rapid regulation of nuclear proteins by rapamycin-induced translocation in fission yeast.
abstract: Genetic analysis of protein function requires a rapid means of inactivating the gene under study. Typically, this exploits temperature-sensitive mutations or promoter shut-off techniques. We report the adaptation to Schizosaccharomyces pombe of the anchor-away technique, originally designed in budding yeast by Laemmli lab. This method relies on a rapamycin-mediated interaction between the FRB- and FKBP12-binding domains to relocalize nuclear proteins of interest to the cytoplasm. We demonstrate a rapid nuclear depletion of abundant proteins as proof of principle.
authors.affiliation: Molecular and Computational Biology, University of Southern California, Los Angeles, CA, USA.
authors.forename: Lin
authors.initials: L
authors.lastname: Ding
authors.forename: Dana
authors.initials: D
authors.lastname: Laor
authors.forename: Ronit
authors.initials: R
authors.lastname: Weisman
authors.forename: Susan L
authors.initials: SL
authors.lastname: Forsburg
mesh_terms: D003433:Crosses, Genetic; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D008856:Microscopy, Fluorescence; D064148:Minichromosome Maintenance Complex Component 4 / Q000502:physiology*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012568:Schizosaccharomyces / Q000502:physiology*; D029702:Schizosaccharomyces pombe Proteins / Q000502:physiology*; D020123:Sirolimus / Q000494:pharmacology*; D022061:Tacrolimus Binding Protein 1A / Q000502:physiology*; D014170:Transformation, Genetic
chemical_list: D004271:DNA, Fungal; C480031:Rad11 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D064148:Minichromosome Maintenance Complex Component 4; C078490:mcm4 protein, S pombe; D022061:Tacrolimus Binding Protein 1A; D020123:Sirolimus
keywords: FKBP; FRB; Schizosaccharomyces pombe; anchor-away; nuclear proteins; rapamycin
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