My journey to DNA repair

Abstract

I completed my medical studies at the Karolinska Institute in Stockholm but have always been devoted to basic research. My longstanding interest is to understand fundamental DNA repair mechanisms in the fields of cancer therapy, inherited human genetic disorders and ancient DNA. I initially measured DNA decay, including rates of base loss and cytosine deamination. I have discovered several important DNA repair proteins and determined their mechanisms of action. The discovery of uracil-DNA glycosylase defined a new category of repair enzymes with each specialized for different types of DNA damage. The base excision repair pathway was first reconstituted with human proteins in my group. Cell-free analysis for mammalian nucleotide excision repair of DNA was also developed in my laboratory. I found multiple distinct DNA ligases in mammalian cells, and led the first genetic and biochemical work on DNA ligases I, III and IV. I discovered the mammalian exonucleases DNase III (TREX1) and IV (FEN1). Interestingly, expression of TREX1 was altered in some human autoimmune diseases. I also showed that the mutagenic DNA adduct O(6)-methylguanine (O(6)mG) is repaired without removing the guanine from DNA, identifying a surprising mechanism by which the methyl group is transferred to a residue in the repair protein itself. A further novel process of DNA repair discovered by my research group is the action of AlkB as an iron-dependent enzyme carrying out oxidative demethylation.

Figure 1

Overview of mechanistic models for enzymatic reactions A. (I) DNA glycosylases catalyze the cleavage of base-sugar bonds; (II) AP endonucleases incise double-stranded DNA at base-free sugar-phosphate residues; (III) FEN1 removes overhangs and flaps from DNA and (IV) eukaryotic DNA ligases ligate DNA ends. B. O⁶-methylguanine-DNA methyltransferase (MGMT) transfers irreversibly a promutagenic methyl group from alkylated DNA to a specific cysteine residue in the transferase itself. C. DNA dioxygenases remove certain cytotoxic methyl groups from alkylated base residues by oxidative demethylation in the presence of iron and oxoglutarate. D. FTO and ALKBH5 demethylate RNA m⁶A as a novel epigenetic marker in α-ketoglutarate (α-KG) and Fe²⁺-dependent manner. E. TREX1 is a 3′ to 5′ exonuclease with preference for single-stranded DNA.