Measuring macroautophagy in S. cerevisiae: autophagic body accumulation and total protein turnover.
Macroautophagy has been implicated in various physiological functions and severe human diseases. Accordingly, there is a high interest in determining macroautophagy both qualitatively and quantitatively. In this chapter we discuss how macroautophagy can be followed morphologically in the yeast Saccharomyces cerevisiae using light microscopy. To quantitatively measure macroautophagy, we further present two protocols for the determination of total protein turnover.
Prick
Tanja
T
Georg-August-University, Center of Biochemistry and Molecular Cell Biology, Goettingen, Germany.
Thumm
Michael
M
D000818:Animals; D001343:Autophagy / Q000502:physiology*; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D008853:Microscopy / Q000379:methods*; D010588:Phagosomes / Q000378:metabolism / Q000648:ultrastructure; D011726:Pyridinium Compounds / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D011868:Radioisotopes / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism / Q000648:ultrastructure
C092350:FM 4-64; D005456:Fluorescent Dyes; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D011868:Radioisotopes; D029701:Saccharomyces cerevisiae Proteins