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Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator

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 populati...

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Autores principales: Jung, Jennifer, Nayak, Arnab, Schaeffer, Véronique, Starzetz, Tatjana, Kirsch, Achim K, Müller, Stefan, Dikic, Ivan, Mittelbronn, Michel, Behrends, Christian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5323046/
https://www.ncbi.nlm.nih.gov/pubmed/28195531
http://dx.doi.org/10.7554/eLife.23063
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author Jung, Jennifer
Nayak, Arnab
Schaeffer, Véronique
Starzetz, Tatjana
Kirsch, Achim K
Müller, Stefan
Dikic, Ivan
Mittelbronn, Michel
Behrends, Christian
author_facet Jung, Jennifer
Nayak, Arnab
Schaeffer, Véronique
Starzetz, Tatjana
Kirsch, Achim K
Müller, Stefan
Dikic, Ivan
Mittelbronn, Michel
Behrends, Christian
author_sort Jung, Jennifer
collection PubMed
description 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. DOI: http://dx.doi.org/10.7554/eLife.23063.001
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spelling pubmed-53230462017-02-27 Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator Jung, Jennifer Nayak, Arnab Schaeffer, Véronique Starzetz, Tatjana Kirsch, Achim K Müller, Stefan Dikic, Ivan Mittelbronn, Michel Behrends, Christian eLife Biochemistry 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. DOI: http://dx.doi.org/10.7554/eLife.23063.001 eLife Sciences Publications, Ltd 2017-02-14 /pmc/articles/PMC5323046/ /pubmed/28195531 http://dx.doi.org/10.7554/eLife.23063 Text en © 2017, Jung et al http://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Biochemistry
Jung, Jennifer
Nayak, Arnab
Schaeffer, Véronique
Starzetz, Tatjana
Kirsch, Achim K
Müller, Stefan
Dikic, Ivan
Mittelbronn, Michel
Behrends, Christian
Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator
title Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator
title_full Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator
title_fullStr Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator
title_full_unstemmed Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator
title_short Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator
title_sort multiplex image-based autophagy rnai screening identifies smcr8 as ulk1 kinase activity and gene expression regulator
topic Biochemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5323046/
https://www.ncbi.nlm.nih.gov/pubmed/28195531
http://dx.doi.org/10.7554/eLife.23063
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