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

2013/07/09 by Magdalena Morawska, Helle D. Ulrich · 8 citations
Biochemistry, Genetics and Molecular Biology · Agricultural and Biological Sciences · #Ubiquitin and proteasome pathways #Plant Virus Research Studies #Protein Degradation and Inhibitors

paper · pdf · doi:10.1002/yea.2967

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.

Citations

• Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae
• Epitope tagging of yeast genes using a PCR-based strategy: more tags and improved practical routines

Cited by

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• Controllable targeted protein degradation as a promising tool for discovery of novel cellular and developmental mechanisms
• The mitotic STAG3–cohesin complex shapes male germline nucleome
• Inducible tag-free degradation of endogenous proteins with AlissAID and development of a photoactivable inducer
• A proteome-wide yeast degron collection for the dynamic study of protein function
• Significantly reduced, but balanced, rates of mitochondrial fission and fusion are sufficient to maintain the integrity of yeast mitochondrial DNA

Related

• OsTIR1 ^F74G Expression Controls Basal and Auxin-Induced Degradation Rates of the AID System in Yeast
• A proteome-wide yeast degron collection for the dynamic study of protein function
• Inducible tag-free degradation of endogenous proteins with AlissAID and development of a photoactivable inducer
• Epitope tagging of yeast genes using a PCR-based strategy: more tags and improved practical routines
• A versatile toolbox for PCR‐based tagging of yeast genes: new fluorescent proteins, more markers and promoter substitution cassettes