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Monitoring of Iron Depletion-Induced Mitophagy in Pathogenic Yeast

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Mitophagy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1759))

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.

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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

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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.

Author information

Authors and Affiliations

  1. Department of Food Science and Human Nutrition, Faculty of Agriculture, Ryukoku University, 1-5 Yokotani, Seta Oe-cho, Otsu, Shiga, 520-2194, Japan

    Koichi Tanabe

  2. Department of Chemotherapy and Mycoses, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan

    Koichi Tanabe & Minoru Nagi

Authors
  1. Koichi Tanabe
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  2. Minoru Nagi
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Corresponding author

Correspondence to Koichi Tanabe .

Editor information

Editors and Affiliations

  1. Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan

    Nobutaka Hattori

  2. Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan

    Shinji Saiki

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Tanabe, K., Nagi, M. (2017). Monitoring of Iron Depletion-Induced Mitophagy in Pathogenic Yeast. In: Hattori, N., Saiki, S. (eds) Mitophagy. Methods in Molecular Biology, vol 1759. Humana Press, New York, NY. https://doi.org/10.1007/7651_2017_40

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  • DOI: https://doi.org/10.1007/7651_2017_40

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7749-9

  • Online ISBN: 978-1-4939-7750-5

  • eBook Packages: Springer Protocols

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