Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator
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- Jennifer Jung
- Institute of Biochemistry II, Goethe University School of Medicine, Frankfurt, Germany
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- Arnab Nayak
- Institute of Biochemistry II, Goethe University School of Medicine, Frankfurt, Germany
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- Véronique Schaeffer
- Institute of Biochemistry II, Goethe University School of Medicine, Frankfurt, Germany
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- Tatjana Starzetz
- Neurological Institute, Goethe University, Frankfurt, Germany
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- Achim K Kirsch
- PerkinElmer, Inc., Hamburg, Germany
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- Stefan Müller
- Institute of Biochemistry II, Goethe University School of Medicine, Frankfurt, Germany
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- Ivan Dikic
- Institute of Biochemistry II, Goethe University School of Medicine, Frankfurt, Germany
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- Michel Mittelbronn
- Neurological Institute, Goethe University, Frankfurt, Germany
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- Christian Behrends
- Institute of Biochemistry II, Goethe University School of Medicine, Frankfurt, Germany
説明
<jats:p>Autophagy is an intracellular recycling and degradation pathway that depends on membrane trafficking. Rab GTPases are central for autophagy but their regulation especially through the activity of Rab GEFs remains largely elusive. We employed a RNAi screen simultaneously monitoring different populations of autophagosomes and identified 34 out of 186 Rab GTPase, GAP and GEF family members as potential autophagy regulators, amongst them SMCR8. SMCR8 uses overlapping binding regions to associate with C9ORF72 or with a C9ORF72-ULK1 kinase complex holo-assembly, which function in maturation and formation of autophagosomes, respectively. While focusing on the role of SMCR8 during autophagy initiation, we found that kinase activity and gene expression of ULK1 are increased upon SMCR8 depletion. The latter phenotype involved association of SMCR8 with the ULK1 gene locus. Global mRNA expression analysis revealed that SMCR8 regulates transcription of several other autophagy genes including WIPI2. Collectively, we established SMCR8 as multifaceted negative autophagy regulator.</jats:p>
収録刊行物
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- eLife
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eLife 6 1-, 2017-02-14
eLife Sciences Publications, Ltd
