title: An expanded tool kit for the auxin-inducible degron system in budding yeast.
abstract: Fusion of inducible degradation signals, so-called degrons, to cellular proteins is an elegant method of controlling protein levels in vivo. Recently, a degron system relying on the plant hormone auxin has been described for use in yeast and vertebrate cells. We now report the construction of a series of vectors that significantly enhance the versatility of this auxin-inducible degron (AID) system in Saccharomyces cerevisiae. We have minimized the size of the degron and appended a series of additional epitope tags, allowing detection by commercial antibodies or fluorescence microscopy. The vectors are compatible with PCR-based genomic tagging strategies, allow for C- or N-terminal fusion of the degron, and provide a range of selection markers. Application to a series of yeast proteins, including essential replication factors, provides evidence for a general usefulness of the system.
authors.affiliation: Cancer Research UK London Research Institute, Clare Hall Laboratories, South Mimms, UK.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Morawska
authors.forename: Helle D
authors.initials: HD
authors.lastname: Ulrich
mesh_terms: D015966:Gene Expression Regulation, Fungal / Q000187:drug effects*; D005822:Genetic Vectors; D005827:Genetics, Microbial / Q000379:methods*; D007210:Indoleacetic Acids / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*; D009172:Mycology / Q000379:methods; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D007210:Indoleacetic Acids; D029701:Saccharomyces cerevisiae Proteins
keywords: auxin; budding yeast; degron; protein degradation; protein stability