Glyceraldehyde-3-phosphate Dehydrogenase (GAPDH) Aggregation Causes Mitochondrial Dysfunction during Oxidative Stress-induced Cell Death.
Glycolytic glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a multifunctional protein that also mediates cell death under oxidative stress. We reported previously that the active-site cysteine (Cys-152) of GAPDH plays an essential role in oxidative stress-induced aggregation of GAPDH associated with cell death, and a C152A-GAPDH mutant rescues nitric oxide (NO)-induced cell death by interfering with the aggregation of wild type (WT)-GAPDH. However, the detailed mechanism underlying GAPDH aggregate-induced cell death remains elusive. Here we report that NO-induced GAPDH aggregation specifically causes mitochondrial dysfunction. First, we observed a correlation between NO-induced GAPDH aggregation and mitochondrial dysfunction, when GAPDH aggregation occurred at mitochondria in SH-SY5Y cells. In isolated mitochondria, aggregates of WT-GAPDH directly induced mitochondrial swelling and depolarization, whereas mixtures containing aggregates of C152A-GAPDH reduced mitochondrial dysfunction. Additionally, treatment with cyclosporin A improved WT-GAPDH aggregate-induced swelling and depolarization. In doxycycline-inducible SH-SY5Y cells, overexpression of WT-GAPDH augmented NO-induced mitochondrial dysfunction and increased mitochondrial GAPDH aggregation, whereas induced overexpression of C152A-GAPDH significantly suppressed mitochondrial impairment. Further, NO-induced cytochromecrelease into the cytosol and nuclear translocation of apoptosis-inducing factor from mitochondria were both augmented in cells overexpressing WT-GAPDH but ameliorated in C152A-GAPDH-overexpressing cells. Interestingly, GAPDH aggregates induced necrotic cell death via a permeability transition pore (PTP) opening. The expression of either WT- or C152A-GAPDH did not affect other cell death pathways associated with protein aggregation, such as proteasome inhibition, gene expression induced by endoplasmic reticulum stress, or autophagy. Collectively, these results suggest that NO-induced GAPDH aggregation specifically induces mitochondrial dysfunction via PTP opening, leading to cell death.
From the Laboratory of Veterinary Pharmacology, hnakajima@vet.osakafu-u.ac.jp.
Hidemitsu
0000-0002-0441-1929
H
Nakajima
From the Laboratory of Veterinary Pharmacology.
Masanori
M
Itakura
From the Laboratory of Veterinary Pharmacology.
Takeya
T
Kubo
From the Laboratory of Veterinary Pharmacology.
Akihiro
A
Kaneshige
the Division of Applied Life Sciences, and.
Naoki
N
Harada
the Laboratory of Veterinary Pathology, Graduate School of Life and Environmental Science, Osaka Prefecture University, Izumisano, Osaka 5988531, Japan.
Takeshi
T
Izawa
From the Laboratory of Veterinary Pharmacology.
Yasu-Taka
YT
Azuma
the Laboratory of Veterinary Pathology, Graduate School of Life and Environmental Science, Osaka Prefecture University, Izumisano, Osaka 5988531, Japan.
Mitsuru
M
Kuwamura
the Division of Applied Life Sciences, and.
Ryouichi
R
Yamaji
From the Laboratory of Veterinary Pharmacology.
Tadayoshi
T
Takeuchi
D051033:Apoptosis Inducing Factor / Q000378:metabolism; D016923:Cell Death; D045744:Cell Line, Tumor; D045304:Cytochromes c / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism*; D006801:Humans; D008928:Mitochondria / Q000378:metabolism / Q000473:pathology*; D033681:Mitochondrial Membrane Transport Proteins / Q000378:metabolism; D000083162:Mitochondrial Permeability Transition Pore; D009569:Nitric Oxide / Q000378:metabolism*; D018384:Oxidative Stress; D066329:Protein Aggregates
D051033:Apoptosis Inducing Factor; D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D066329:Protein Aggregates; D009569:Nitric Oxide; D045304:Cytochromes c; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
amyloid; cell death; mitochondria; nitric oxide; protein aggregation