The effect of p53-RNAi and p53 knockout on human 8-oxoguanine DNA glycosylase (hOgg1) activity.
Recent evidence indicates that in vitro p53 augments base excision repair (BER) activities in mammalian cells. To understand the role of p53 in BER, we analyzed the repair activity of hOgg1 in isogenic cell lines HCT116p53+/+ and HCT116p53-/-. We found that hOgg1 activity was significantly decreased in HCT116p53-/- cells as compared with HCT116p53+/+ cells, indicating a functional role for p53 in the regulation of hOGG1. Using gel-shift assays, we showed that p53 binds to its putative cis-elements within the hOGG1 promoter. In addition we demonstrated that supplementing p53 in HCT116p53-/- cells enhanced the transcription of hOGG1. To further strengthen our findings, we used p53-RNAi to study the effects of decreased p53 levels on hOgg1 activity. We observed that p53-RNAi resulted in decreased hOGG1 expression both at the mRNA and protein levels. This decrease in hOGG1 expression was associated with reduced cell viability upon oxidative damage and reduced hOgg1 activity as evidenced by the 8-oxoG incision assay. Taken together, our results indicate that loss of p53 function can lead to decreased hOgg1 repair activity.
Chatterjee
Aditi
A
Department of Otolaryngology-Head and Neck Surgery, Head and Neck Cancer Research Division, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2196, USA.
Mambo
Elizabeth
E
Osada
Motonobu
M
Upadhyay
Sunil
S
Sidransky
David
D
D004249:DNA Damage; D045647:DNA Glycosylases / Q000235:genetics / Q000378:metabolism*; D004260:DNA Repair; D015536:Down-Regulation; D017353:Gene Deletion; D015971:Gene Expression Regulation, Enzymologic; D045325:HCT116 Cells; D006801:Humans; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D034622:RNA Interference; D016159:Tumor Suppressor Protein p53 / Q000172:deficiency* / Q000235:genetics* / Q000378:metabolism
D016159:Tumor Suppressor Protein p53; D045647:DNA Glycosylases; C479058:oxoguanine glycosylase 1, human