Skip to main content
SpringerLink
Log in

Methods for Studying Mitophagy in Yeast

  • Protocol
  • First Online:
Autophagy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1880))

Abstract

Under some experimental conditions, eukaryotic cells, from yeast to man, will digest a portion of their mitochondrial cohort through an autophagic process termed mitophagy. In humans, defects in mitophagy have been proposed to play a causative role in a number of late-onset degenerative diseases such as Parkinson’s disease and type II diabetes. As a consequence the study of mitophagy, as a quality control process in eukaryotic cells, has become an increasingly important focus in contemporary cell biology. When faced with the task of assaying mitophagy in yeast, the experimentalist has at his or her disposal a variety of induction conditions and assay systems to choose from. Here, we survey several well-established protocols for inducing and monitoring mitophagy in the yeast Saccharomyces cerevisiae and discuss their relative merits, limitations, and potential pitfalls.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 189.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 249.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Ashford TP, Porter KR (1962) Cytoplasmic components in hepatic cell lysosomes. J Cell Biol 12:198–202

    Article  CAS  Google Scholar 

  2. Deter RL, Baudhuin P, De Duve C (1967) Participation of lysosomes in cellular autophagy induced in rat liver by glucagon. J Cell Biol 35(2):C11–C16

    Article  CAS  Google Scholar 

  3. Lemasters JJ (2005) Selective mitochondrial autophagy, or mitophagy, as a targeted defense against oxidative stress, mitochondrial dysfunction, and aging. Rejuvenation Res 8(1):3–5

    Article  CAS  Google Scholar 

  4. Braell WA, Balch WE, Dobbertin DC, Rothman JE (1984) The glycoprotein that is transported between successive compartments of the Golgi in a cell-free system resides in stacks of cisternae. Cell 39(3 Pt 2):511–524

    Article  CAS  Google Scholar 

  5. Kanki T, Klionsky DJ (2008) Mitophagy in yeast occurs through a selective mechanism. J Biol Chem 283(47):32386–32393

    Article  CAS  Google Scholar 

  6. Okamoto K, Kondo-Okamoto N, Ohsumi Y (2009) Mitochondria-anchored receptor Atg32 mediates degradation of mitochondria via selective autophagy. Dev Cell 17(1):87–97

    Article  CAS  Google Scholar 

  7. Journo D, Mor A, Abeliovich H (2009) Aup1-mediated regulation of Rtg3 during mitophagy. J Biol Chem 284(51):35885–35895

    Article  CAS  Google Scholar 

  8. Kanki T et al (2009) A genomic screen for yeast mutants defective in selective mitochondria autophagy. Mol Biol Cell 20(22):4730–4738

    Article  CAS  Google Scholar 

  9. De Deken RH (1966) The Crabtree effect: a regulatory system in yeast. J Gen Microbiol 44(2):149–156

    Article  Google Scholar 

  10. Hagman A, Säll T, Piškur J (2014) Analysis of the yeast short-term Crabtree effect and its origin. FEBS J 281(21):4805–4814

    Article  CAS  Google Scholar 

  11. Tal R, Winter G, Ecker N, Klionsky DJ, Abeliovich H (2007) Aup1p, a yeast mitochondrial protein phosphatase homolog, is required for efficient stationary phase mitophagy and cell survival. J Biol Chem 282(8):5617–5624

    Article  CAS  Google Scholar 

  12. Heo JM et al (2010) A stress-responsive system for mitochondrial protein degradation. Mol Cell 40(3):465–480

    Article  CAS  Google Scholar 

  13. Vida TA, Emr SD (1995) A new vital stain for visualizing vacuolar membrane dynamics and endocytosis in yeast. J Cell Biol 128(5):779–792

    Article  CAS  Google Scholar 

  14. Kanki T, Kang D, Klionsky DJ (2009) Monitoring mitophagy in yeast: the Om45-GFP processing assay. Autophagy 5(8):1186–1189

    Article  CAS  Google Scholar 

  15. Kissova I, Deffieu M, Manon S, Camougrand N (2004) Uth1p is involved in the autophagic degradation of mitochondria. J Biol Chem 279(37):39068–39074

    Article  CAS  Google Scholar 

  16. Rosado CJ, Mijaljica D, Hatzinisiriou I, Prescott M, Devenish RJ (2008) Rosella: a fluorescent pH-biosensor for reporting vacuolar turnover of cytosol and organelles in yeast. Autophagy 4(2):205–213

    Article  CAS  Google Scholar 

  17. Noda T, Matsuura A, Wada Y, Ohsumi Y (1995) Novel system for monitoring autophagy in the yeast Saccharomyces cerevisiae. Biochem Biophys Res Commun 210(1):126–132

    Article  CAS  Google Scholar 

  18. Klionsky DJ, Emr SD (1990) A new class of lysosomal/vacuolar protein sorting signals. J Biol Chem 265(10):5349–5352

    CAS  PubMed  Google Scholar 

  19. Abeliovich H, Zarei M, Rigbolt KT, Youle RJ, Dengjel J (2013) Involvement of mitochondrial dynamics in the segregation of mitochondrial matrix proteins during stationary phase mitophagy. Nat Commun 4:2789

    Article  Google Scholar 

  20. Marangoni AG (2003) Enzyme kinetics: a modern approach. John Wiley & Sons, Hoboken, NJ

    Google Scholar 

  21. Kaneko Y, Toh-e A, Banno I, Oshima Y (1989) Molecular characterization of a specific p-nitrophenylphosphatase gene, PHO13, and its mapping by chromosome fragmentation in Saccharomyces cerevisiae. Mol Gen Genet 220(1):133–139

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biochemistry, Food Science and Nutrition, Hebrew University of Jerusalem, Rehovot, Israel

    Panagiota Kolitsida & Hagai Abeliovich

Authors
  1. Panagiota Kolitsida
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hagai Abeliovich
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hagai Abeliovich .

Editor information

Editors and Affiliations

  1. Signalling Programme, Babraham Institute, Cambridge, UK

    Nicholas Ktistakis

  2. Signalling Programme, Babraham Institute, Cambridge, UK

    Oliver Florey

Rights and permissions

Reprints and permissions

Copyright information

© 2019 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Kolitsida, P., Abeliovich, H. (2019). Methods for Studying Mitophagy in Yeast. In: Ktistakis, N., Florey, O. (eds) Autophagy. Methods in Molecular Biology, vol 1880. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8873-0_44

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8873-0_44

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8872-3

  • Online ISBN: 978-1-4939-8873-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation