PINK1 autophosphorylation upon membrane potential dissipation is essential for Parkin recruitment to damaged mitochondria.
Dysfunction of PINK1, a mitochondrial Ser/Thr kinase, causes familial Parkinson's disease (PD). Recent studies have revealed that PINK1 is rapidly degraded in healthy mitochondria but accumulates on the membrane potential (ΔΨm)-deficient mitochondria, where it recruits another familial PD gene product, Parkin, to ubiquitylate the damaged mitochondria. Despite extensive study, the mechanism underlying the homeostatic control of PINK1 remains unknown. Here we report that PINK1 is autophosphorylated following a decrease in ΔΨm and that most disease-relevant mutations hinder this event. Mass spectrometric and mutational analyses demonstrate that PINK1 autophosphorylation occurs at Ser228 and Ser402, residues that are structurally clustered together. Importantly, Ala mutation of these sites abolishes autophosphorylation of PINK1 and inhibits Parkin recruitment onto depolarized mitochondria, whereas Asp (phosphorylation-mimic) mutation promotes mitochondrial localization of Parkin even though autophosphorylation was still compromised. We propose that autophosphorylation of Ser228 and Ser402 in PINK1 is essential for efficient mitochondrial localization of Parkin.
Laboratory of Protein Metabolism, Tokyo Metropolitan Institute of Medical Science, Setagaya-ku, 156-8506, Japan.
Kei
K
Okatsu
Toshihiko
T
Oka
Masahiro
M
Iguchi
Kenji
K
Imamura
Hidetaka
H
Kosako
Naoki
N
Tani
Mayumi
M
Kimura
Etsu
E
Go
Fumika
F
Koyano
Manabu
M
Funayama
Kahori
K
Shiba-Fukushima
Shigeto
S
Sato
Hideaki
H
Shimizu
Yuko
Y
Fukunaga
Hisaaki
H
Taniguchi
Masaaki
M
Komatsu
Nobutaka
N
Hattori
Katsuyoshi
K
Mihara
Keiji
K
Tanaka
Noriyuki
N
Matsuda
D000595:Amino Acid Sequence; D000818:Animals; D006367:HeLa Cells; D006801:Humans; D008564:Membrane Potentials; D051379:Mice; D008928:Mitochondria / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010300:Parkinson Disease / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D016415:Sequence Alignment; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000099202:PTEN-Induced Putative Kinase
D011494:Protein Kinases; D044767:Ubiquitin-Protein Ligases; D000099202:PTEN-Induced Putative Kinase; C111567:parkin protein