title: Synthetic quantitative array technology identifies the Ubp3-Bre5 deubiquitinase complex as a negative regulator of mitophagy.
abstract: Mitophagy is crucial to ensuring mitochondrial quality control. However, the molecular mechanism and regulation of mitophagy are still not fully understood. Here, we developed a quantitative methodology termed synthetic quantitative array (SQA) technology, which allowed us to perform a genome-wide screen for modulators of rapamycin-induced mitophagy in S. cerevisiae. SQA technology can be easily employed for other enzyme-based reporter systems and widely applied in yeast research. We identified 86 positive and 10 negative regulators of mitophagy. Moreover, SQA-based analysis of non-selective autophagy revealed that 63 of these regulators are specific for mitophagy and 33 regulate autophagy in general. The Ubp3-Bre5 deubiquitination complex was found to inhibit mitophagy but, conversely, to promote other types of autophagy, including ribophagy. This complex translocates dynamically to mitochondria upon induction of mitophagy. These findings point to a role of ubiquitination in mitophagy in yeast and suggest a reciprocal regulation of distinct autophagy pathways.
authors.affiliation: Mitochondrial Biology, Buchmann Institute for Molecular Life Sciences and Zentrum für Molekulare Medizin, Goethe University, Max-von-Laue-Straße 15, 60438 Frankfurt am Main, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Müller
authors.affiliation: Institut für Molekulare Biowissenschaften, Goethe University, Max-von-Laue-Straße 9, 60438 Frankfurt am Main, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Kötter
authors.affiliation: Mitochondrial Biology, Buchmann Institute for Molecular Life Sciences and Zentrum für Molekulare Medizin, Goethe University, Max-von-Laue-Straße 15, 60438 Frankfurt am Main, Germany; Institute of Biochemistry and Molecular Biology I, Medical Faculty, Heinrich Heine University, Universitätsstraße 1, 40225 Düsseldorf, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Behrendt
authors.affiliation: Mitochondrial Biology, Buchmann Institute for Molecular Life Sciences and Zentrum für Molekulare Medizin, Goethe University, Max-von-Laue-Straße 15, 60438 Frankfurt am Main, Germany.
authors.forename: Elena
authors.initials: E
authors.lastname: Walter
authors.affiliation: Mitochondrial Biology, Buchmann Institute for Molecular Life Sciences and Zentrum für Molekulare Medizin, Goethe University, Max-von-Laue-Straße 15, 60438 Frankfurt am Main, Germany.
authors.forename: Christian Q
authors.initials: CQ
authors.lastname: Scheckhuber
authors.affiliation: Institut für Molekulare Biowissenschaften, Goethe University, Max-von-Laue-Straße 9, 60438 Frankfurt am Main, Germany.
authors.forename: Karl-Dieter
authors.initials: KD
authors.lastname: Entian
authors.affiliation: Mitochondrial Biology, Buchmann Institute for Molecular Life Sciences and Zentrum für Molekulare Medizin, Goethe University, Max-von-Laue-Straße 15, 60438 Frankfurt am Main, Germany; Institute of Biochemistry and Molecular Biology I, Medical Faculty, Heinrich Heine University, Universitätsstraße 1, 40225 Düsseldorf, Germany. Electronic address: reichert@hhu.de.
authors.forename: Andreas S
authors.initials: AS
authors.lastname: Reichert
mesh_terms: D001343:Autophagy / Q000187:drug effects; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D016681:Genome, Fungal; D008928:Mitochondria / Q000378:metabolism*; D063306:Mitophagy / Q000187:drug effects; D016296:Mutagenesis; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D020123:Sirolimus / Q000494:pharmacology; D054875:Ubiquitination
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D010450:Endopeptidases; C503886:Bre5 protein, S cerevisiae; C101111:UBP3 protein, S cerevisiae; D020123:Sirolimus