Nix is critical to two distinct phases of mitophagy, reactive oxygen species-mediated autophagy induction and Parkin-ubiquitin-p62-mediated mitochondrial priming

Wen Xing Ding, Hong Min Ni, Min Li, Yong Liao, Xiaoyun Chen, Donna B. Stolz, Gerald W. Dorn, Xiao-Ming Yin

Research output: Contribution to journalArticle

322 Citations (Scopus)

Abstract

Damaged mitochondria can be eliminated by autophagy, i.e. mitophagy, which is important for cellular homeostasis and cell survival. Despite the fact that a number of factors have been found to be important for mitophagy in mammalian cells, their individual roles in the process had not been clearly defined. Parkin is a ubiquitin-protein isopeptide ligase able to translocate to the mitochondria that are to be removed. We showed here in a chemical hypoxia model of mitophagy induced by an uncoupler, carbonyl cyanide m-chlorophenylhydrazone (CCCP) that Parkin translocation resulted in mitochondrial ubiquitination and p62 recruitment to the mitochondria. Small inhibitory RNA-mediated knockdown of p62 significantly diminished mitochondrial recognition by the autophagy machinery and the subsequent elimination. Thus Parkin, ubiquitin, and p62 function in preparing mitochondria for mitophagy, here referred to as mitochondrial priming. However, these molecules were not required for the induction of autophagy machinery. Neither Parkin nor p62 seemed to affect autophagy induction by CCCP. Instead, we found that Nix was required for the autophagy induction. Nix promoted CCCP-induced mitochondrial depolarization and reactive oxygen species generation, which inhibited mTOR signaling and activated autophagy. Nix also contributed to mitochondrial priming by controlling the mitochondrial translocation of Parkin, although reactive oxygen species generation was not involved in this step. Deletion of the C-terminal membrane targeting sequence but not mutations in the BH3 domain disabled Nix for these functions. Our work thus distinguished the molecular events responsible for the different phases of mitophagy and placed Nix upstream of the events.

Original languageEnglish
Pages (from-to)27879-27890
Number of pages12
JournalJournal of Biological Chemistry
Volume285
Issue number36
DOIs
StatePublished - Sep 3 2010

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Mitochondrial Degradation
Mitochondria
Autophagy
Ubiquitin
Reactive Oxygen Species
Machinery
Cells
Ubiquitin-Protein Ligases
Depolarization
Chemical Models
Ubiquitination
RNA
Membranes
Molecules
Cell Survival
Homeostasis
mesoxalonitrile
Mutation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

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Nix is critical to two distinct phases of mitophagy, reactive oxygen species-mediated autophagy induction and Parkin-ubiquitin-p62-mediated mitochondrial priming. / Ding, Wen Xing; Ni, Hong Min; Li, Min; Liao, Yong; Chen, Xiaoyun; Stolz, Donna B.; Dorn, Gerald W.; Yin, Xiao-Ming.

In: Journal of Biological Chemistry, Vol. 285, No. 36, 03.09.2010, p. 27879-27890.

Research output: Contribution to journalArticle

Ding, Wen Xing ; Ni, Hong Min ; Li, Min ; Liao, Yong ; Chen, Xiaoyun ; Stolz, Donna B. ; Dorn, Gerald W. ; Yin, Xiao-Ming. / Nix is critical to two distinct phases of mitophagy, reactive oxygen species-mediated autophagy induction and Parkin-ubiquitin-p62-mediated mitochondrial priming. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 36. pp. 27879-27890.
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