TY - JOUR
T1 - Oxidation of mitochondrial deoxynucleotide pools by exposure to sodium nitroprusside induces cell death
AU - Ichikawa, Junji
AU - Tsuchimoto, Daisuke
AU - Oka, Sugako
AU - Ohno, Mizuki
AU - Furuichi, Masato
AU - Sakumi, Kunihiko
AU - Nakabeppu, Yusaku
N1 - Funding Information:
We are grateful to Dr. K. Mihara for his generous gift of ABCme cDNA, Dr. Mitsuo Iida for providing us with the opportunity to conduct this study, and Drs. W. Campbell and D. Yoshimura for useful comments on this manuscript. We thank N. Adachi for HPLC MS/MS analysis, and M. Sasaki and M. Ohtsu in the Laboratory for Technical Support of our institute for their expertise in performing the EM and DNA sequence analyses. This work was supported by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (grant numbers: 16012248, 17014070, 18013038), and the Japan Society for the Promotion of Science (grant numbers: 16390119, 18300124).
PY - 2008/3/1
Y1 - 2008/3/1
N2 - Human MutT homolog (hMTH1) hydrolyzes oxidized purine nucleoside triphosphates to monophosphates, thereby avoiding incorporation of such oxidized purines into DNA or RNA. We examined whether hMTH1 prevents cellular dysfunction induced by sodium nitroprusside, a spontaneous NO donor. Exposure to sodium nitroprusside caused an 8-oxoguanine (8-oxoG) buildup in DNA of proliferating MTH1-null cells which underwent mitochondrial degeneration and subsequently died. Quiescent MTH1-null cells also died with 8-oxoG buildup but only when the buildup affected mitochondrial and not nuclear DNA. In both proliferative and quiescent conditions, the accumulation of 8-oxoG in DNA and cell death was effectively prevented by hMTH1. Knockdown of MUTYH in quiescent MTH1-null cells significantly prevented the cell death, suggesting that 8-oxoG incorporated into mitochondrial DNA is a main cause of this form of cell death. To verify this possibility, an artificially modified hMTH1, namely mTP-EGFP-hMTH1, which localizes exclusively in mitochondria, was expressed in MTH1-null cells. mTP-EGFP-hMTH1 selectively prevented buildup of 8-oxoG in mitochondrial but not nuclear DNA after exposure of proliferating cells to sodium nitroprusside, and also efficiently prevented cell death. We thus concluded that exposure of cells to sodium nitroprusside causes oxidation of mitochondrial deoxynucleotide pools, and that buildup of oxidized bases in mitochondrial DNA initiates cell death.
AB - Human MutT homolog (hMTH1) hydrolyzes oxidized purine nucleoside triphosphates to monophosphates, thereby avoiding incorporation of such oxidized purines into DNA or RNA. We examined whether hMTH1 prevents cellular dysfunction induced by sodium nitroprusside, a spontaneous NO donor. Exposure to sodium nitroprusside caused an 8-oxoguanine (8-oxoG) buildup in DNA of proliferating MTH1-null cells which underwent mitochondrial degeneration and subsequently died. Quiescent MTH1-null cells also died with 8-oxoG buildup but only when the buildup affected mitochondrial and not nuclear DNA. In both proliferative and quiescent conditions, the accumulation of 8-oxoG in DNA and cell death was effectively prevented by hMTH1. Knockdown of MUTYH in quiescent MTH1-null cells significantly prevented the cell death, suggesting that 8-oxoG incorporated into mitochondrial DNA is a main cause of this form of cell death. To verify this possibility, an artificially modified hMTH1, namely mTP-EGFP-hMTH1, which localizes exclusively in mitochondria, was expressed in MTH1-null cells. mTP-EGFP-hMTH1 selectively prevented buildup of 8-oxoG in mitochondrial but not nuclear DNA after exposure of proliferating cells to sodium nitroprusside, and also efficiently prevented cell death. We thus concluded that exposure of cells to sodium nitroprusside causes oxidation of mitochondrial deoxynucleotide pools, and that buildup of oxidized bases in mitochondrial DNA initiates cell death.
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U2 - 10.1016/j.dnarep.2007.11.007
DO - 10.1016/j.dnarep.2007.11.007
M3 - Article
C2 - 18155646
AN - SCOPUS:39149100596
VL - 7
SP - 418
EP - 430
JO - DNA Repair
JF - DNA Repair
SN - 1568-7864
IS - 3
ER -