Miro phosphorylation sites regulate Parkin recruitment and mitochondrial motility.
The PTEN-induced putative kinase 1 (PINK1)/Parkin pathway can tag damaged mitochondria and trigger their degradation by mitophagy. Before the onset of mitophagy, the pathway blocks mitochondrial motility by causing Miro degradation. PINK1 activates Parkin by phosphorylating both Parkin and ubiquitin. PINK1, however, has other mitochondrial substrates, including Miro (also called RhoT1 and -2), although the significance of those substrates is less clear. We show that mimicking PINK1 phosphorylation of Miro on S156 promoted the interaction of Parkin with Miro, stimulated Miro ubiquitination and degradation, recruited Parkin to the mitochondria, and via Parkin arrested axonal transport of mitochondria. Although Miro S156E promoted Parkin recruitment it was insufficient to trigger mitophagy in the absence of broader PINK1 action. In contrast, mimicking phosphorylation of Miro on T298/T299 inhibited PINK1-induced Miro ubiquitination, Parkin recruitment, and Parkin-dependent mitochondrial arrest. The effects of the T298E/T299E phosphomimetic were dominant over S156E substitution. We propose that the status of Miro phosphorylation influences the decision to undergo Parkin-dependent mitochondrial arrest, which, in the context of PINK1 action on other substrates, can restrict mitochondrial dynamics before mitophagy.
F. M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, MA 02115; Department of Neurobiology, Harvard Medical School, Harvard University, Cambridge, MA 02138.
Evgeny
E
Shlevkov
F. M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, MA 02115; Department of Neurobiology, Harvard Medical School, Harvard University, Cambridge, MA 02138.
Tal
T
Kramer
Ann Romney Center for Neurological Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115.
Jason
J
Schapansky
Ann Romney Center for Neurological Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115.
Matthew J
MJ
LaVoie
F. M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, MA 02115; Department of Neurobiology, Harvard Medical School, Harvard University, Cambridge, MA 02138; Thomas.Schwarz@childrens.harvard.edu.
Thomas L
TL
Schwarz
D019943:Amino Acid Substitution; D000596:Amino Acids / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001370:Axonal Transport; D017930:Genes, Reporter; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008928:Mitochondria / Q000378:metabolism*; D063154:Mitochondrial Dynamics; D063306:Mitophagy / Q000235:genetics; D009154:Mutation; D010766:Phosphorylation; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism; D059748:Proteolysis; D017966:Pyramidal Cells / Q000378:metabolism; D051381:Rats; D055647:Rats, Transgenic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*; D054875:Ubiquitination; D020741:rho GTP-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000099202:PTEN-Induced Putative Kinase
D000596:Amino Acids; D011494:Protein Kinases; D011993:Recombinant Fusion Proteins; D044767:Ubiquitin-Protein Ligases; D020741:rho GTP-Binding Proteins; D000099202:PTEN-Induced Putative Kinase; C111567:parkin protein
Miro; PINK1; Parkin; mitochondrial transport; mitophagy
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