Alkyladenine DNA glycosylase deficiency uncouples alkylation-induced strand break generation from PARP-1 activation and glycolysis inhibition

• Published in: Scientific reports Vol. 10; no. 1; p. 2209
• Main Authors: Alhumaydhi, Fahad A., de O. Lopes, Debora, Bordin, Diana L., Aljohani, Abdullah S. M., Lloyd, Cameron B., McNicholas, Michael D., Milano, Larissa, Charlier, Clara F., Villela, Izabel, Henriques, João Antonio P., Plant, Kathryn E., Elliott, Ruan M., Meira, Lisiane B.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 10.02.2020; Nature Publishing Group
• Subjects: 631/45; 631/45/147; Acrylamides - pharmacology; Adenosine diphosphate; Alkylation; Animals; Base excision repair; Cells, Cultured; Cytokines - antagonists & inhibitors; Cytokines - metabolism; Cytotoxicity; Deoxyribonucleic acid; DNA; DNA Breaks - drug effects; DNA damage; DNA glycosylase; DNA Glycosylases - deficiency; DNA Glycosylases - genetics; DNA Repair; Enzymes; Fibroblasts; Glycolysis; Glycolysis - drug effects; Humanities and Social Sciences; Intermediates; Metabolic response; Metabolism; Methyl methanesulfonate; Methyl Methanesulfonate - pharmacology; Mice; Mice, Knockout; Mitochondria; multidisciplinary; NAD; NAD - metabolism; Nicotinamide; Nicotinamide phosphoribosyltransferase; Nicotinamide Phosphoribosyltransferase - antagonists & inhibitors; Nicotinamide Phosphoribosyltransferase - metabolism; Phosphoribosyltransferase; Piperidines - pharmacology; Poly (ADP-Ribose) Polymerase-1 - metabolism; Poly(ADP-ribose) polymerase; Primary Cell Culture; Ribose; Science; Science (multidisciplinary)
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-59072-6

DNA alkylation damage is repaired by base excision repair (BER) initiated by alkyladenine DNA glycosylase (AAG). Despite its role in DNA repair, AAG-initiated BER promotes cytotoxicity in a process dependent on poly (ADP-ribose) polymerase-1 (PARP-1); a NAD + -consuming enzyme activated by strand break intermediates of the AAG-initiated repair process. Importantly, PARP-1 activation has been previously linked to impaired glycolysis and mitochondrial dysfunction. However, whether alkylation affects cellular metabolism in the absence of AAG-mediated BER initiation is unclear. To address this question, we temporally profiled repair and metabolism in wild-type and Aag −/− cells treated with the alkylating agent methyl methanesulfonate (MMS). We show that, although Aag −/− cells display similar levels of alkylation-induced DNA breaks as wild type, PARP-1 activation is undetectable in AAG-deficient cells. Accordingly, Aag −/− cells are protected from MMS-induced NAD + depletion and glycolysis inhibition. MMS-induced mitochondrial dysfunction, however, is AAG-independent. Furthermore, treatment with FK866, a selective inhibitor of the NAD + salvage pathway enzyme nicotinamide phosphoribosyltransferase (NAMPT), synergizes with MMS to induce cytotoxicity and Aag −/− cells are resistant to this combination FK866 and MMS treatment. Thus, AAG plays an important role in the metabolic response to alkylation that could be exploited in the treatment of conditions associated with NAD + dysregulation.