Immunofluorescent localization of the murine 8-oxoguanine DNA glycosylase (mOGG1) in cells growing under normal and nutrient deprivation conditions.
OGG1 is a major DNA glycosylase in mammalian cells, participating in the repair of 7,8-dihydro-8-oxoguanine (8-oxoguanine, 8-oxoG), the most abundant known DNA lesion induced by endogenous reactive oxygen species in aerobic organisms. 8-oxoG is therefore often used as a marker for oxidative DNA damage. In this study, polyclonal and monoclonal antibodies were raised against the purified wild-type recombinant murine 8-oxoG DNA glycosylase (mOGG1) protein and their specificity against the native enzyme and the SDS-denatured mOGG1 polypeptide were characterized. Specific antibodies directed against the purified wild-type recombinant mOGG1 were used to localize in situ this DNA repair enzyme in established cell lines (HeLa cells, NIH3T3 fibroblasts) as well as in primary culture mouse embryo fibroblasts growing under either normal or oxidative stress conditions. Results from these studies showed that mOGG1 is localized to the nucleus and the cytoplasm of mammalian cells in culture. However, mOGG1 levels increase and primarily redistribute to the nucleus and its peripheral cytoplasm in cells exposed to oxidative stress conditions. Immunofluorescent localization results reported in this study suggest that susceptibility to oxidative DNA damage varies among mammalian tissue culture cells and that mOGG1 appears to redistribute once mOGG1 cell copy number increases in response to oxidative DNA damage.
Conlon
Kimberly A
KA
Department of Pharmacological Sciences, School of Medicine, University Hospital and Medical Center, State University of New York at Stony Brook, Stony Brook, NY 11794-8651, USA.
Zharkov
Dmitry O
DO
Berrios
Miguel
M
D000818:Animals; D002455:Cell Division; D002467:Cell Nucleus / Q000201:enzymology*; D003593:Cytoplasm / Q000201:enzymology*; D004249:DNA Damage; D045647:DNA Glycosylases / Q000378:metabolism*; D004260:DNA Repair; D004622:Embryo, Mammalian / Q000166:cytology / Q000201:enzymology; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells / Q000166:cytology / Q000201:enzymology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D041681:NIH 3T3 Cells / Q000166:cytology / Q000201:enzymology; D018384:Oxidative Stress; D011817:Rabbits; D011994:Recombinant Proteins
D007074:Immunoglobulin G; D011994:Recombinant Proteins; D045647:DNA Glycosylases; C485990:Ogg1 protein, mouse