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Parkin interacts with Ambra1 to induce mitophagy

Cindy Van Humbeeck, Tom Cornelissen, Hilde Hofkens, Wim Mandemakers, Kris Gevaert UGent, Bart De Strooper and Wim Vandenberghe (2011) JOURNAL OF NEUROSCIENCE. 31(28). p.10249-10261
abstract
Mutations in the gene encoding Parkin are a major cause of recessive Parkinson's disease. Recent work has shown that Parkin translocates from the cytosol to depolarized mitochondria and induces their autophagic removal (mitophagy). However, the molecular mechanisms underlying Parkin-mediated mitophagy are poorly understood. Here, we investigated whether Parkin interacts with autophagy-regulating proteins. We purified Parkin and associated proteins from HEK293 cells using tandem affinity purification and identified the Parkin interactors using mass spectrometry. We identified the autophagy-promoting protein Ambra1 (activating molecule in Beclin1-regulated autophagy) as a Parkin interactor. Ambra1 activates autophagy in the CNS by stimulating the activity of the class III phosphatidylinositol 3-kinase (PI3K) complex that is essential for the formation of new phagophores. We found Ambra1, like Parkin, to be widely expressed in adult mouse brain, including midbrain dopaminergic neurons. Endogenous Parkin and Ambra1 coimmunoprecipitated from HEK293 cells, SH-SY5Y cells, and adult mouse brain. We found no evidence for ubiquitination of Ambra1 by Parkin. The interaction of endogenous Parkin and Ambra1 strongly increased during prolonged mitochondrial depolarization. Ambra1 was not required for Parkin translocation to depolarized mitochondria but was critically important for subsequent mitochondrial clearance. In particular, Ambra1 was recruited to perinuclear clusters of depolarized mitochondria and activated class III PI3K in their immediate vicinity. These data identify interaction of Parkin with Ambra1 as a key mechanism for induction of the final clearance step of Parkin-mediated mitophagy.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
P62/SQSTM1, ACTIVATION, MEMBRANE, PINK1, MICE, DISEASE, AUTOPHAGY, NERVOUS-SYSTEM, MITOCHONDRIAL-FUNCTION, UBIQUITIN-PROTEIN LIGASE
journal title
JOURNAL OF NEUROSCIENCE
J. Neurosci.
volume
31
issue
28
pages
10249 - 10261
Web of Science type
Article
Web of Science id
000292699600016
JCR category
NEUROSCIENCES
JCR impact factor
7.115 (2011)
JCR rank
19/242 (2011)
JCR quartile
1 (2011)
ISSN
0270-6474
DOI
10.1523/JNEUROSCI.1917-11.2011
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
1863778
handle
http://hdl.handle.net/1854/LU-1863778
date created
2011-07-29 16:30:25
date last changed
2016-12-19 15:41:56
@article{1863778,
  abstract     = {Mutations in the gene encoding Parkin are a major cause of recessive Parkinson's disease. Recent work has shown that Parkin translocates from the cytosol to depolarized mitochondria and induces their autophagic removal (mitophagy). However, the molecular mechanisms underlying Parkin-mediated mitophagy are poorly understood. Here, we investigated whether Parkin interacts with autophagy-regulating proteins. We purified Parkin and associated proteins from HEK293 cells using tandem affinity purification and identified the Parkin interactors using mass spectrometry. We identified the autophagy-promoting protein Ambra1 (activating molecule in Beclin1-regulated autophagy) as a Parkin interactor. Ambra1 activates autophagy in the CNS by stimulating the activity of the class III phosphatidylinositol 3-kinase (PI3K) complex that is essential for the formation of new phagophores. We found Ambra1, like Parkin, to be widely expressed in adult mouse brain, including midbrain dopaminergic neurons. Endogenous Parkin and Ambra1 coimmunoprecipitated from HEK293 cells, SH-SY5Y cells, and adult mouse brain. We found no evidence for ubiquitination of Ambra1 by Parkin. The interaction of endogenous Parkin and Ambra1 strongly increased during prolonged mitochondrial depolarization. Ambra1 was not required for Parkin translocation to depolarized mitochondria but was critically important for subsequent mitochondrial clearance. In particular, Ambra1 was recruited to perinuclear clusters of depolarized mitochondria and activated class III PI3K in their immediate vicinity. These data identify interaction of Parkin with Ambra1 as a key mechanism for induction of the final clearance step of Parkin-mediated mitophagy.},
  author       = {Van Humbeeck, Cindy and Cornelissen, Tom and Hofkens, Hilde and Mandemakers, Wim and Gevaert, Kris and De Strooper, Bart and Vandenberghe, Wim},
  issn         = {0270-6474},
  journal      = {JOURNAL OF NEUROSCIENCE},
  keyword      = {P62/SQSTM1,ACTIVATION,MEMBRANE,PINK1,MICE,DISEASE,AUTOPHAGY,NERVOUS-SYSTEM,MITOCHONDRIAL-FUNCTION,UBIQUITIN-PROTEIN LIGASE},
  language     = {eng},
  number       = {28},
  pages        = {10249--10261},
  title        = {Parkin interacts with Ambra1 to induce mitophagy},
  url          = {http://dx.doi.org/10.1523/JNEUROSCI.1917-11.2011},
  volume       = {31},
  year         = {2011},
}

Chicago
Van Humbeeck, Cindy, Tom Cornelissen, Hilde Hofkens, Wim Mandemakers, Kris Gevaert, Bart De Strooper, and Wim Vandenberghe. 2011. “Parkin Interacts with Ambra1 to Induce Mitophagy.” Journal of Neuroscience 31 (28): 10249–10261.
APA
Van Humbeeck, C., Cornelissen, T., Hofkens, H., Mandemakers, W., Gevaert, K., De Strooper, B., & Vandenberghe, W. (2011). Parkin interacts with Ambra1 to induce mitophagy. JOURNAL OF NEUROSCIENCE, 31(28), 10249–10261.
Vancouver
1.
Van Humbeeck C, Cornelissen T, Hofkens H, Mandemakers W, Gevaert K, De Strooper B, et al. Parkin interacts with Ambra1 to induce mitophagy. JOURNAL OF NEUROSCIENCE. 2011;31(28):10249–61.
MLA
Van Humbeeck, Cindy, Tom Cornelissen, Hilde Hofkens, et al. “Parkin Interacts with Ambra1 to Induce Mitophagy.” JOURNAL OF NEUROSCIENCE 31.28 (2011): 10249–10261. Print.