Probing aggrephagy using chemically-induced protein aggregates

Anne F. J. Janssen, Eugene A. Katrukha, Wendy van Straaten, Pauline Verlhac, Fulvio Reggiori, Lukas C. Kapitein*

*Bijbehorende auteur voor dit werk

OnderzoeksoutputAcademicpeer review

14 Citaten (Scopus)
285 Downloads (Pure)

Samenvatting

Selective types of autophagy mediate the clearance of specific cellular components and are essential to maintain cellular homeostasis. However, tools to directly induce and monitor such pathways are limited. Here we introduce the PIM (particles induced by multimerization) assay as a tool for the study of aggrephagy, the autophagic clearance of aggregates. The assay uses an inducible multimerization module to assemble protein clusters, which upon induction recruit ubiquitin, p62, and LC3 before being delivered to lysosomes. Moreover, use of a dual fluorescent tag allows for the direct observation of cluster delivery to the lysosome. Using flow cytometry and fluorescence microscopy, we show that delivery to the lysosome is partially dependent on p62 and ATG7. This assay will help in elucidating the spatiotemporal dynamics and control mechanisms underlying aggregate clearance by the autophagy-lysosomal system.

Originele taal-2English
Artikelnummer4245
Pagina's (van-tot)4245
Aantal pagina's10
TijdschriftNature Communications
Volume9
Nummer van het tijdschrift1
DOI's
StatusPublished - 12-okt-2018

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