title: Conditional nuclear import and export of yeast proteins using a chemical inducer of dimerization.
abstract: In eukaryotes, reversible shuttling between the nucleus and cytoplasm is an important regulatory mechanism, particularly for many kinases and transcription factors. Inspired by the natural system, we recently developed a technology to control protein position in budding yeast using a chemical inducer of dimerization (CID). In this method, a nuclear export or localization signal is reversibly appended to a protein of interest by the CID, which effectively places its subcellular location under direct control of the chemical stimulus. Here, we explicitly tested the ability of this system to direct the nucleocytoplasmic transport of a panel of 16 representative kinases and transcription factors. From this set, we found that 12 targets (75%) are susceptible to re-positioning, suggesting that this method might be applicable to a range of targets. Interestingly, the four proteins that resisted mislocalization (Fun20p, Hcm1p, Pho4p, and Ste12p) are known to engage in a large number of protein-protein contacts. We suspect that, for these highly connected targets, the strength of the chemical signal may be insufficient to drive mislocalization and that proteins with relatively few partners might be most amenable to this approach. Collectively, these studies provide a necessary framework for the design of large-scale applications.
authors.affiliation: Department of Pathology, University of Michigan, Ann Arbor, MI 48109-2216, USA.
authors.forename: Srikanth
authors.initials: S
authors.lastname: Patury
authors.forename: Prasanthi
authors.initials: P
authors.lastname: Geda
authors.forename: Craig J
authors.initials: CJ
authors.lastname: Dobry
authors.forename: Anuj
authors.initials: A
authors.lastname: Kumar
authors.forename: Jason E
authors.initials: JE
authors.lastname: Gestwicki
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D032701:Biomimetics; D002467:Cell Nucleus / Q000378:metabolism*; D025141:Drug Resistance, Fungal; D005800:Genes, Fungal / Q000235:genetics; D010641:Phenotype; D010770:Phosphotransferases / Q000737:chemistry / Q000378:metabolism; D055503:Protein Multimerization / Q000187:drug effects*; D020836:Protein Structure, Quaternary / Q000187:drug effects; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism*; D020123:Sirolimus / Q000494:pharmacology*; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D010770:Phosphotransferases; D020123:Sirolimus