title: Targeted protein depletion in Saccharomyces cerevisiae by activation of a bidirectional degron.
abstract: BACKGROUND
  Tools for in vivo manipulation of protein abundance or activity are highly beneficial for life science research. Protein stability can be efficiently controlled by conditional degrons, which induce target protein degradation at restrictive conditions.
  
  
  RESULTS
  We used the yeast Saccharomyces cerevisiae for development of a conditional, bidirectional degron to control protein stability, which can be fused to the target protein N-terminally, C-terminally or placed internally. Activation of the degron is achieved by cleavage with the tobacco etch virus (TEV) protease, resulting in quick proteolysis of the target protein. We found similar degradation rates of soluble substrates using destabilization by the N- or C-degron. C-terminal tagging of essential yeast proteins with the bidirectional degron resulted in deletion-like phenotypes at non-permissive conditions. Developmental process-specific mutants were created by N- or C-terminal tagging of essential proteins with the bidirectional degron in combination with sporulation-specific production of the TEV protease.
  
  
  CONCLUSIONS
  We developed a system to influence protein abundance and activity genetically, which can be used to create conditional mutants, to regulate the fate of single protein domains or to design artificial regulatory circuits. Thus, this method enhances the toolbox to manipulate proteins in systems biology approaches considerably.
authors.affiliation: Department of Genetics, Philipps-Universität Marburg, Germany.
authors.forename: Marc
authors.initials: M
authors.lastname: Jungbluth
authors.forename: Christian
authors.initials: C
authors.lastname: Renicke
authors.forename: Christof
authors.initials: C
authors.lastname: Taxis
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids; D000818:Animals; D010450:Endopeptidases / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D009955:Ornithine Decarboxylase / Q000737:chemistry / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013997:Time Factors
chemical_list: D000596:Amino Acids; D005656:Fungal Proteins; D010450:Endopeptidases; C099488:TEV protease; D009955:Ornithine Decarboxylase