Monitoring mitophagy in yeast

Nadine Camougrand, Ingrid Kissova, Benedicte Salin, Rodney James Devenish

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

57 Citations (Scopus)

Abstract

Cellular degradative processes including proteasomal and vacuolar/lysosomal autophagic degradation, as well as the activity of proteases (both cytosolic and mitochondrial), provide for a continuous turnover of damaged and obsolete macromolecules and organelles. Mitochondria are essential for oxidative energy production in aerobic eukaryotic cells, where they are also required for multiple biosynthetic pathways to take place. Mitochondrial homeostasis also plays a crucial role in aging and programmed cell death, and recent data have suggested that mitochondrial degradation is a strictly regulated process. A recent study has shown that in yeast cells subjected to nitrogen starvation, degradation of mitochondria by autophagy occurs by both a selective process (termed mitophagy) and a nonselective process. This chapter provides an overview of the techniques that enable the study of mitophagy. Fluorescent proteins targeted to mitochondria can be used to follow mitochondrial sequestration within vacuoles. Degradation of mitochondria can be assayed using a mitochondrially targeted alkaline phosphatase (ALP) reporter test in which the delivery of mitochondrial N-terminal truncated Pho8Delta60 to the vacuole results from mitophagy. Degradation of mitochondrial proteins can also be followed by Western immunoblot analyses. Finally, electron microscopy observations permit the discrimination between selective mitophagy and nonselective mitochondrial degradation.
Original languageEnglish
Title of host publicationMethods in Enzymology: Autophagy: Lower Eukaryotes and Non-Mammalian Systems
EditorsDJ Klionsky
Place of PublicationUSA
PublisherAcademic Press
Pages89 - 107
Number of pages19
Edition451
ISBN (Print)9780123745484
Publication statusPublished - 2009

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