A fluorescent tool set for yeast Atg proteins

Autophagy. 2015;11(6):954-60. doi: 10.1080/15548627.2015.1040971.

Abstract

Fluorescence microscopy of live cells is instrumental in deciphering the molecular details of autophagy. To facilitate the routine examination of yeast Atg proteins under diverse conditions, here we provide a comprehensive tool set, including (1) plasmids for the expression of GFP chimeras at endogenous levels for most Atg proteins, (2) RFP-Atg8 constructs with improved properties as a PAS marker, and (3) plasmids for the complementation of common yeast auxotrophic markers. We hope that the availability of this tool set will further accelerate yeast autophagy research.

Keywords: Atg, autophagy related; C,G,R,YFP, cyan, green, red and yellow fluorescent protein; Cvt, cytoplasm-to-vacuole targeting; DsRed eExpress 2; PAS, phagophore assembly site; Vps, vacuolar protein sorting.; autophagy; auxotroph; fluorescent protein; mKO, monomeric Kusabira Orange; pseudo-monomer; starter kit; yeast.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autophagy / physiology*
  • Autophagy-Related Protein 8 Family
  • Autophagy-Related Proteins
  • Green Fluorescent Proteins / metabolism
  • Microscopy, Fluorescence*
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism*
  • Phagosomes / metabolism
  • Protein Kinases / genetics
  • Protein Kinases / metabolism*
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Vacuoles / metabolism

Substances

  • ATG8 protein, S cerevisiae
  • Autophagy-Related Protein 8 Family
  • Autophagy-Related Proteins
  • Microtubule-Associated Proteins
  • Saccharomyces cerevisiae Proteins
  • Green Fluorescent Proteins
  • Protein Kinases
  • ATG1 protein, S cerevisiae