MiT/TFE transcription factors are activated during mitophagy downstream of Parkin and Atg5

J Cell Biol. 2015 Aug 3;210(3):435-50. doi: 10.1083/jcb.201501002.

Abstract

The kinase PINK1 and ubiquitin ligase Parkin can regulate the selective elimination of damaged mitochondria through autophagy (mitophagy). Because of the demand on lysosomal function by mitophagy, we investigated a role for the transcription factor EB (TFEB), a master regulator of lysosomal biogenesis, in this process. We show that during mitophagy TFEB translocates to the nucleus and displays transcriptional activity in a PINK1- and Parkin-dependent manner. MITF and TFE3, homologues of TFEB belonging to the same microphthalmia/transcription factor E (MiT/TFE) family, are similarly regulated during mitophagy. Unlike TFEB translocation after starvation-induced mammalian target of rapamycin complex 1 inhibition, Parkin-mediated TFEB relocalization required Atg9A and Atg5 activity. However, constitutively active Rag guanosine triphosphatases prevented TFEB translocation during mitophagy, suggesting cross talk between these two MiT/TFE activation pathways. Analysis of clustered regularly interspaced short palindromic repeats-generated TFEB/MITF/TFE3/TFEC single, double, and triple knockout cell lines revealed that these proteins partly facilitate Parkin-mediated mitochondrial clearance. These results illuminate a pathway leading to MiT/TFE transcription factor activation, distinct from starvation-induced autophagy, which occurs during mitophagy.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • 14-3-3 Proteins / metabolism
  • Active Transport, Cell Nucleus
  • Autophagy / genetics
  • Autophagy / physiology*
  • Autophagy-Related Protein 5
  • Autophagy-Related Proteins
  • Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / genetics
  • Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / metabolism*
  • Cell Line, Tumor
  • Gene Knockout Techniques
  • HEK293 Cells
  • HeLa Cells
  • Homeodomain Proteins / metabolism
  • Humans
  • Lysosomes / metabolism
  • Mechanistic Target of Rapamycin Complex 1
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Microphthalmia-Associated Transcription Factor / genetics
  • Microtubule-Associated Proteins / genetics
  • Microtubule-Associated Proteins / metabolism*
  • Mitochondria / metabolism
  • Mitochondrial Proteins / genetics
  • Mitophagy / genetics
  • Mitophagy / physiology*
  • Multiprotein Complexes / antagonists & inhibitors
  • Phagosomes / metabolism
  • Protein Kinases / genetics
  • RNA Interference
  • RNA, Small Interfering
  • TOR Serine-Threonine Kinases / antagonists & inhibitors
  • Ubiquitin-Protein Ligases / genetics
  • Ubiquitin-Protein Ligases / metabolism*
  • Vesicular Transport Proteins / metabolism
  • rab GTP-Binding Proteins / genetics
  • rab7 GTP-Binding Proteins

Substances

  • 14-3-3 Proteins
  • ATG5 protein, human
  • Atg9a protein, human
  • Autophagy-Related Protein 5
  • Autophagy-Related Proteins
  • Basic Helix-Loop-Helix Leucine Zipper Transcription Factors
  • FIS1 protein, human
  • Homeodomain Proteins
  • MITF protein, human
  • Membrane Proteins
  • Microphthalmia-Associated Transcription Factor
  • Microtubule-Associated Proteins
  • Mitochondrial Proteins
  • Multiprotein Complexes
  • RNA, Small Interfering
  • TFE3 protein, human
  • TFEB protein, human
  • Vesicular Transport Proteins
  • rab7 GTP-Binding Proteins
  • RAG-1 protein
  • Ubiquitin-Protein Ligases
  • parkin protein
  • Protein Kinases
  • Mechanistic Target of Rapamycin Complex 1
  • PTEN-induced putative kinase
  • TOR Serine-Threonine Kinases
  • rab GTP-Binding Proteins