Human Base Excision Repair Creates a Bias Toward −1 Frameshift Mutations

• Published in: The Journal of biological chemistry 2010-08, Vol.285 (33), p.25203-25212
• Main Authors: Lyons, Derek M., O'Brien, Patrick J.
• Format: Article
• Language: English
• Subjects: Alkyladenine DNA Glycosylase; Base Excision Repair; DNA and Chromosomes; DNA Damage; DNA Damage - genetics; DNA Damage - physiology; DNA Enzymes; DNA Glycosylases - genetics; DNA Glycosylases - metabolism; DNA Repair; DNA Repair - genetics; DNA Repair - physiology; Enzyme Mechanisms; Enzymology; Frameshift; Frameshift Mutation - genetics; Glycosylase; HeLa Cells; Humans; Models, Genetic; Nucleic Acid Enzymology; Uracil DNA Glycosylase; Uracil-DNA Glycosidase - genetics; Uracil-DNA Glycosidase - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M110.118596

Frameshift mutations are particularly deleterious to protein function and play a prominent role in carcinogenesis. Most commonly these mutations involve the insertion or omission of a single nucleotide by a DNA polymerase that slips on a damaged or undamaged template. The mismatch DNA repair pathway can repair these nascent polymerase errors. However, overexpression of enzymes of the base excision repair (BER) pathway is known to increase the frequency of frameshift mutations suggesting competition between these pathways. We have examined the fate of DNA containing single nucleotide bulges in human cell extracts and discovered that several deaminated or alkylated nucleotides are efficiently removed by BER. Because single nucleotide bulges are more highly exposed we anticipate that they would be highly susceptible to spontaneous DNA damage. As a model for this, we have shown that chloroacetaldehyde reacts more than 18-fold faster with an A-bulge than with a stable A·T base pair to create alkylated DNA adducts that can be removed by alkyladenine DNA glycosylase. Reconstitution of the BER pathway using purified components establishes that bulged DNA is efficiently processed. Single nucleotide deletion is predicted to repair +1 frameshift events, but to make −1 frameshift events permanent. Therefore, these findings suggest an additional factor contributing to the bias toward deletion mutations.