Suppression of uracil-DNA glycosylase induces neuronal apoptosis.
A chronic imbalance in DNA precursors, caused by one-carbon metabolism impairment, can result in a deficiency of DNA repair and increased DNA damage. Although indirect evidence suggests that DNA damage plays a role in neuronal apoptosis and in the pathogenesis of neurodegenerative disorders, the underlying mechanisms are poorly understood. In particular, very little is known about the role of base excision repair of misincorporated uracil in neuronal survival. To test the hypothesis that repair of DNA damage associated with uracil misincorporation is critical for neuronal survival, we employed an antisense (AS) oligonucleotide directed against uracil-DNA glycosylase encoded by the UNG gene to deplete UNG in cultured rat hippocampal neurons. AS, but not a scrambled control oligonucleotide, induced apoptosis, which was associated with DNA damage analyzed by comet assay and up-regulation of p53. UNG mRNA and protein levels were decreased within 30 min and were undetectable within 6-9 h of exposure to the UNG AS oligonucleotide. Whereas UNG expression is significantly higher in proliferating as compared with nonproliferating cells, such as neurons, the levels of UNG mRNA were increased in brains of cystathionine beta-synthase knockout mice, a model for hyperhomocysteinemia, suggesting that one-carbon metabolism impairment and uracil misincorporation can induce the up-regulation of UNG expression.
Kruman
Inna I
II
Sun Health Research Institute, Sun City, Arizona 85351, USA. inna.kruman@sunhealth.org
Schwartz
Elena
E
Kruman
Yuri
Y
Cutler
Roy G
RG
Zhu
Xiaoxiang
X
Greig
Nigel H
NH
Mattson
Mark P
MP
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D016923:Cell Death; D002455:Cell Division; D002478:Cells, Cultured; D004249:DNA Damage; D045647:DNA Glycosylases / Q000187:drug effects / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006624:Hippocampus / Q000166:cytology / Q000196:embryology; D007700:Kinetics; D009474:Neurons / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D014498:Uracil / Q000378:metabolism
D016376:Oligonucleotides, Antisense; D014498:Uracil; D045647:DNA Glycosylases