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Monitoring of Iron Depletion-Induced Mitophagy in Pathogenic Yeast

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Part of the Methods in Molecular Biology book series

Abstract

Mitophagy, which is the degradation of mitochondria via selective autophagic machinery, is thought to be involved in regulating the mass and function of mitochondria. Methods for detection of mitophagy have been reported for several fungal cells including some budding yeast, methylotrophic yeast, and filamentous fungi. Mitophagy in Saccharomyces cerevisiae is activated under nitrogen-poor conditions; however, the regulatory mechanism of mitophagy in most fungi has not been elucidated. Here we describe methods to monitor mitophagy in the pathogenic yeast Candida glabrata under iron-depleted conditions but not under nitrogen starvation. This observation may provide some clues to elucidate the physiological roles of mitophagy in eukaryotes.

Keywords

Atg32 Autophagy Candida Iron Mitochondria Mitophagy Pathogenicity Yeast 

Abbreviations

DHFR

Dihydrofolate reductase

ECL

Enhanced chemiluminescence

GFP

Green fluorescent protein

OD

Optical density

PMSF

Phenylmethane sulfonyl fluoride

ROS

Reactive oxygen species

SD

Synthetic glucose medium

TTBS

Tris-buffered saline containing Tween 20

WT

Wild-type

YPD

Yeast extract peptone dextrose

Notes

Acknowledgement

We thank Koji and Noriko Okamoto (Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan) for providing yeast strains and plasmids. This research was supported by JSPS KAKENHI Grant Numbers 26790428 (KT), 24590540 (HN), and 26860296 (MN). This work was partly supported by a grant from the Ministry of Health, Labour and Welfare of Japan (H26-shinkoujitsuyouka-ippan-010). The authors would like to thank Enago (www.enago.jp) for the English language review.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Food Science and Human Nutrition, Faculty of AgricultureRyukoku UniversityOtsuJapan
  2. 2.Department of Chemotherapy and MycosesNational Institute of Infectious DiseasesTokyoJapan

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