Universität zu Köln

Xiong, Qiuhong, Fischer, Sarah ORCID: 0000-0001-6227-3950, Karow, Malte, Mueller, Rolf, Messling, Susanne and Eichinger, Ludwig ORCID: 0000-0003-1594-6117 (2018). ATG16 mediates the autophagic degradation of the 19S proteasomal subunits PSMD1 and PSMD2. Eur. J. Cell Biol., 97 (8). S. 523 - 533. MUNICH: ELSEVIER GMBH. ISSN 1618-1298

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Abstract

Autophagy and the ubiquitin proteasome system are the two major cellular processes for protein and organelle recycling and clearance in eukaryotic cells. Evidence is accumulating that these two pathways are interrelated through adaptor proteins. Here, we found that PSMD1 and PSMD2, both components of the 19S regulatory particle of the proteasome, directly interact with Dictyostelium discoideum autophagy 16 (ATG16), a core autophagosomal protein. ATG16 is composed of an N-terminal domain, which is responsible for homo-dimerization and binding to ATG5 and a C-terminal beta-propeller structure. Deletion analysis of ATG16 showed that the N-terminal half of ATG16 interacted directly only with PSMD1, while the C-terminal half interacted with both, PSMD1 and PSMD2. RFP-tagged PSMD1 as well as PSMD2 were enriched in large puncta, reminiscent of autophagosomes, in wild-type cells. These puncta were absent in at;g16(-) and atg9(-)/16(-) cells and weaker and less frequent in atg9(-) cells, showing that ATG16 was crucial and the autophagic process important for their formation. Co-expression of ATG16-GFP or GFP-ATG8a(LC3) with RFP-PSMD1 or RFP-PSMD2, respectively, in atg16(-) or wild-type cells revealed many instances of co-localization, suggesting that RFP-PSMD1 or RFP-PSMD2 positive puncta constitute autophagosomes. LysoTracker labeling and a proteolytic cleavage assay confirmed that PSMD1 and PSMD2 were present in lysosomes in wild-type cells. In vivo, ATG16 is required for their enrichment in ATG8a positive puncta, which mature into autolysosomes. We propose that ATG16 links autophagy and the ubiquitin proteasome system.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Xiong, Qiuhong UNSPECIFIED UNSPECIFIED UNSPECIFIED Fischer, Sarah UNSPECIFIED orcid.org/0000-0001-6227-3950 UNSPECIFIED Karow, Malte UNSPECIFIED UNSPECIFIED UNSPECIFIED Mueller, Rolf UNSPECIFIED UNSPECIFIED UNSPECIFIED Messling, Susanne UNSPECIFIED UNSPECIFIED UNSPECIFIED Eichinger, Ludwig UNSPECIFIED orcid.org/0000-0003-1594-6117 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-166347
DOI:	10.1016/j.ejcb.2018.09.002
Journal or Publication Title:	Eur. J. Cell Biol.
Volume:	97
Number:	8
Page Range:	S. 523 - 533
Date:	2018
Publisher:	ELSEVIER GMBH
Place of Publication:	MUNICH
ISSN:	1618-1298
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language MOLD DICTYOSTELIUM-DISCOIDEUM; SELECTIVE AUTOPHAGY; WD-REPEAT; PROTEIN-DEGRADATION; MEMBRANE; COMPLEX; BINDING; YEAST; OLIGOMERIZATION; RECRUITMENT Multiple languages Cell Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/16634