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Parkin drives pS65-Ub turnover independently of canonical autophagy in Drosophila.

Published version
Peer-reviewed

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Authors

Sanchez-Martinez, Alvaro  ORCID logo  https://orcid.org/0000-0002-2728-6251
Terriente-Felix, Ana 
Chen, Po-Lin 

Abstract

Parkinson's disease-related proteins, PINK1 and Parkin, act in a common pathway to maintain mitochondrial quality control. While the PINK1-Parkin pathway can promote autophagic mitochondrial turnover (mitophagy) following mitochondrial toxification in cell culture, alternative quality control pathways are suggested. To analyse the mechanisms by which the PINK1-Parkin pathway operates in vivo, we developed methods to detect Ser65-phosphorylated ubiquitin (pS65-Ub) in Drosophila. Exposure to the oxidant paraquat led to robust, Pink1-dependent pS65-Ub production, while pS65-Ub accumulates in unstimulated parkin-null flies, consistent with blocked degradation. Additionally, we show that pS65-Ub specifically accumulates on disrupted mitochondria in vivo. Depletion of the core autophagy proteins Atg1, Atg5 and Atg8a did not cause pS65-Ub accumulation to the same extent as loss of parkin, and overexpression of parkin promoted turnover of both basal and paraquat-induced pS65-Ub in an Atg5-null background. Thus, we have established that pS65-Ub immunodetection can be used to analyse Pink1-Parkin function in vivo as an alternative to reporter constructs. Moreover, our findings suggest that the Pink1-Parkin pathway can promote mitochondrial turnover independently of canonical autophagy in vivo.

Description

Funder: Gates Cambridge Trust; Id: http://dx.doi.org/10.13039/501100005370

Keywords

in vivo, Parkinson's disease, mitochondria, mitophagy, phospho-ubiquitin, Animals, Drosophila, Autophagy, Protein Serine-Threonine Kinases, Drosophila Proteins

Journal Title

EMBO Rep

Conference Name

Journal ISSN

1469-221X
1469-3178

Volume Title

Publisher

Springer Science and Business Media LLC
Sponsorship
Medical Research Council (MC_UU_00015/6)
Medical Research Council (MC_UU_00015/7)