P150glued-associated disorders are caused by activation of intrinsic apoptotic pathway.
Mutations in p150glued cause hereditary motor neuropathy with vocal cord paralysis (HMN7B) and Perry syndrome (PS). Here we show that both overexpression of p150glued mutants and knockdown of endogenous p150glued induce apoptosis. Overexpression of a p150glued plasmid containing either a HMN7B or PS mutation resulted in cytoplasmic p150glued-positive aggregates and was associated with cell death. Cells containing mutant p150glued aggregates underwent apoptosis that was characterized by an increase in cleaved caspase-3- or Annexin V-positive cells and was attenuated by both zVAD-fmk (a pan-caspase inhibitor) application and caspase-3 siRNA knockdown. In addition, overexpression of mutant p150glued decreased mitochondrial membrane potentials and increased levels of translocase of the mitochondrial outer membrane (Tom20) protein, indicating accumulation of damaged mitochondria. Importantly, siRNA knockdown of endogenous p150glued independently induced apoptosis via caspase-8 activation and was not associated with mitochondrial morphological changes. Simultaneous knockdown of endogenous p150glued and overexpression of mutant p150glued had additive apoptosis induction effects. These findings suggest that both p150glued gain-of-toxic-function and loss-of-physiological-function can cause apoptosis and may underlie the pathogenesis of p150glued-associated disorders.
Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan.
Kei-Ichi
K
Ishikawa
Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan.
Shinji
S
Saiki
Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan; Department of Research and Therapeutics for Movement Disorders, Juntendo University School of Medicine, Tokyo, Japan.
Norihiko
N
Furuya
Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan.
Daisuke
D
Yamada
Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan.
Yoko
Y
Imamichi
Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan.
Yuanzhe
Y
Li
Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan.
Sumihiro
S
Kawajiri
Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan.
Hironori
H
Sasaki
Department of Cell Biology and Neuroscience, Juntendo University School of Medicine, Tokyo, Japan.
Masato
M
Koike
Department of Neurology, Fukuoka University School of Medicine, Fukuoka, Japan.
Yoshio
Y
Tsuboi
Department of Neurology, Juntendo University School of Medicine, Tokyo, Japan; Department of Research and Therapeutics for Movement Disorders, Juntendo University School of Medicine, Tokyo, Japan.
Nobutaka
N
Hattori
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics*; D053148:Caspase 3 / Q000378:metabolism*; D053181:Caspase 8 / Q000378:metabolism*; D061945:Caspase Inhibitors / Q000494:pharmacology; D000072159:Dynactin Complex; D006367:HeLa Cells; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism
D000590:Amino Acid Chloromethyl Ketones; D061945:Caspase Inhibitors; D000072159:Dynactin Complex; D008869:Microtubule-Associated Proteins; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D053148:Caspase 3; D053181:Caspase 8