AlkB mystery solved: Oxidative demethylation of N1-methyladenine and N3-methylcytosine adducts by a direct reversal mechanism

All organisms have multiple DNA repair pathways to protect against alkylation-induced mutation and cell death. For nearly two decades, we have known that the Escherichia coli alkB gene product protects against cell killing by S _N 2-alkylating agents, probably through DNA repair. Despite numerous attempts, a specific DNA repair activity could not be assigned to AlkB. Now, a breakthrough in biology and biochemistry, coupled with the discovery of an in silico protein structure, has uncovered a novel direct reversal DNA repair mechanism that is catalyzed by AlkB, namely the oxidative demethylation of N1-methyladenine or N3-methylcytosine DNA lesions. This reaction occurs on both single- and double-stranded DNA, and requires AlkB-bound non-heme Fe ²⁺ , O ₂ and α-ketogluterate to oxidize the offending methyl group. This is followed by the release of succinate, CO ₂ and formaldehyde, and the restoration of undamaged A or C in DNA.