Probing aggrephagy using chemically-induced protein aggregatesJanssen, A. F. J., Katrukha, E. A., van Straaten, W., Verlhac, P., Reggiori, F. & Kapitein, L. C., 12-Oct-2018, In : Nature Communications. 9, 1, p. 4245 10 p., 4245.
Research output: Contribution to journal › Article › Academic › peer-review
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.
|Number of pages||10|
|Publication status||Published - 12-Oct-2018|
- SELECTIVE AUTOPHAGY, CARGO RECOGNITION, DEGRADATION, CELLS, LYSOSOMES, DISEASE, RECEPTORS, SIGNALS, LIGHT