Abstract
Mutations in the PINK1 gene cause autosomal recessive Parkinson's disease. The PINK1 gene encodes a protein kinase that is mitochondrially cleaved to generate two mature isoforms. In addition to its protective role against mitochondrial dysfunction and apoptosis, PINK1 is also known to regulate mitochondrial dynamics acting upstream of the PD-related protein Parkin. Recent data showed that mitochondrial Parkin promotes the autophagic degradation of dysfunctional mitochondria, and that stable PINK1 silencing may have an indirect role in mitophagy activation. Here we report a new interaction between PINK1 and Beclin1, a key pro-autophagic protein already implicated in the pathogenesis of Alzheimer's and Huntington's diseases. Both PINK1 N- and C-terminal are required for the interaction, suggesting that full-length PINK1, and not its cleaved isoforms, interacts with Beclin1. We also demonstrate that PINK1 significantly enhances basal and starvation-induced autophagy, which is reduced by knocking down Beclin1 expression or by inhibiting the Beclin1 partner Vps34. A mutant, PINK1W437X, interaction of which with Beclin1 is largely impaired, lacks the ability to enhance autophagy, whereas this is not observed for PINK1G309D, a mutant with defective kinase activity but unaltered ability to bind Beclin1. These findings identify a new function of PINK1 and further strengthen the link between autophagy and proteins implicated in the neurodegenerative process.
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Abbreviations
- 3MA:
-
3-methyladenine
- aa:
-
amino acid
- ER:
-
endoplasmic reticulum
- FL:
-
full length
- FLIM:
-
Fluorescence Lifetime Imaging Microscopy
- FRET:
-
Fluorescence Resonance Energy Transfer
- LC3:
-
Microtubule-associated protein light chain 3
- Mfn1:
-
Mitofusin 1
- MPTP:
-
1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine
- OMM:
-
outer mitochondrial membrane
- PD:
-
Parkinson's disease
- PI3K:
-
phosphatidylinositol 3′-kinase
- PINK1:
-
PTEN induced putative kinase 1
- TRAP1:
-
TNF receptor-associated protein 1
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Acknowledgements
We are grateful to Dr. Sylvia Lombardo for technical support, Dr. Alessandro Ferraris for statistical analysis support, Prof. David Markie for help in designing the yeast two-hybrid experiment and for providing the pGBD-B vector, Dr. Francesca De Marchi for providing GFP–LC3 (pEGFPC1-LC3) and Prof. Luca Scorrano for providing human Mitofusin1 (pCB6Myc-MFN1) and human WT DRP1 (pcDNA3.1-DRP1) plasmids. This study was supported with grants from Telethon Foundation (GGP07210 to EMV and GC), Italian Ministry of Health (Ricerca Corrente 2008–2009, Ricerca Finalizzata 2004 and 2006 and ex art.56 to BD) and Fondazione Cariplo (to GC). The support of Fondazione Livio Patrizi and Transgenomics is also gratefully acknowledged. The sponsors had no role in study design, data collection and analysis, decision to publish or preparation of the paper.
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Michiorri, S., Gelmetti, V., Giarda, E. et al. The Parkinson-associated protein PINK1 interacts with Beclin1 and promotes autophagy. Cell Death Differ 17, 962–974 (2010). https://doi.org/10.1038/cdd.2009.200
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DOI: https://doi.org/10.1038/cdd.2009.200
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