An expanded tool kit for the auxin-inducible degron system in budding yeast

Yeast. 2013 Sep;30(9):341-51. doi: 10.1002/yea.2967. Epub 2013 Jul 23.

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.

Keywords: auxin; budding yeast; degron; protein degradation; protein stability.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Expression Regulation, Fungal / drug effects*
  • Genetic Vectors
  • Genetics, Microbial / methods*
  • Indoleacetic Acids / metabolism*
  • Molecular Biology / methods*
  • Mycology / methods
  • Saccharomyces cerevisiae / drug effects*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / biosynthesis
  • Saccharomyces cerevisiae Proteins / genetics

Substances

  • Indoleacetic Acids
  • Saccharomyces cerevisiae Proteins