Link between mitochondria and NADPH oxidase 1 isozyme for the sustained production of reactive oxygen species and cell death.
Although mitochondria and the Nox family of NADPH oxidase are major sources of reactive oxygen species (ROS) induced by external stimuli, there is limited information on their functional relationship. This study has shown that serum withdrawal promotes the production of ROS in human 293T cells by stimulating both the mitochondria and Nox1. An analysis of their relationship revealed that the mitochondria respond to serum withdrawal within a few minutes, and the ROS produced by the mitochondria trigger Nox1 action by stimulating phosphoinositide 3-kinase (PI3K) and Rac1. Activation of the PI3K/Rac1/Nox1 pathway was evident 4-8 h after but not earlier than serum withdrawal initiation, and this time lag was found to be required for an additional activator of the pathway, Lyn, to be expressed. Functional analysis suggested that, although the mitochondria contribute to the early (0-4 h) accumulation of ROS, the maintenance of the induced ROS levels to the later (4-8 h) phase required the action of the PI3K/Rac1/Nox1 pathway. Serum withdrawal-treated cells eventually lost their viability, which was reversed by blocking either the mitochondria-dependent induction of ROS using rotenone or KCN or the PI3K/Rac1/Nox1 pathway using the dominant negative mutants or small interfering RNAs. This suggests that mitochondrial ROS are essential but not enough to promote cell death, which requires the sustained accumulation of ROS by the subsequent action of Nox1. Overall, this study shows a signaling link between the mitochondria and Nox1, which is crucial for the sustained accumulation of ROS and cell death in serum withdrawal-induced signaling.
Lee
Seung Bum
SB
Laboratory of Radiation Tumor Physiology, Korea Institute of Radiological and Medical Sciences, 215-4 Gongneung-dong, Nowon-gu, Seoul 139-706, Korea.
Bae
In Hwa
IH
Bae
Yun Soo
YS
Um
Hong-Duck
HD
D016923:Cell Death; D002470:Cell Survival; D006801:Humans; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry*; D000074624:NADPH Oxidase 1; D019255:NADPH Oxidases / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D020033:Protein Isoforms; D021381:Protein Transport; D034741:RNA, Small Interfering / Q000378:metabolism; D017382:Reactive Oxygen Species; D015398:Signal Transduction; D020830:rac1 GTP-Binding Protein / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism
D020033:Protein Isoforms; C107953:RAC1 protein, human; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D009247:NADH, NADPH Oxidoreductases; D000074624:NADPH Oxidase 1; D019255:NADPH Oxidases; C402799:NOX1 protein, human; D019869:Phosphatidylinositol 3-Kinases; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases; D020830:rac1 GTP-Binding Protein