title: The Schizosaccharomyces pombe AlkB homolog Abh1 exhibits AP lyase activity but no demethylase activity.
abstract: 2-Oxoglutarate (2OG) and iron (Fe(II)) dependent dioxygenases catalyze a wide range of biological oxidations, including hydroxylation and demethylation of proteins and nucleic acids. AlkB from Escherichia coli directly reverses certain methyl lesions in DNA, and defines a subfamily of 2OG/Fe(II) dioxygenases that has so far been shown to be involved in both nucleic acid repair and modification. The human genome encodes nine AlkB homologs and the function of most of these is still unknown. The fission yeast Schizosaccharomyces pombe has two AlkB homologs and here we have addressed the function of one of these, Abh1, which appears not to possess a classical AlkB-like repair activity. No enzymatic activity was found toward methylated DNA or etheno adducts, nor was the yeast abh1- mutant sensitive toward alkylating agents. Interestingly, heterologous expression of E. coli AlkB protected the fission yeast cells from alkylation induced cytotoxicity, suggesting that S. pombe lacks systems for efficient repair of lesions that are AlkB substrates. Further, we show that Abh1 possesses an unexpected DNA incision activity at apurinic/apyrimidinic (AP) sites. This AP lyase activity did not depend on 2OG and Fe(II) and was not repressed by dioxygenase inhibitors. Survival and complementation analyses failed to reveal any biological role for AP lyase cleavage by Abh1. It appears that in vitro AP lyase activity can be detected for a number of enzymes belonging to structurally and functionally unrelated families, but the in vivo significance of such activities may be questionable.
authors.affiliation: Department of Microbiology, Oslo University Hospital HF Rikshospitalet, and Centre for Molecular Biology and Neuroscience (CMBN), University of Oslo, Oslo, Norway.
authors.forename: Hanne
authors.initials: H
authors.lastname: Korvald
authors.forename: Pål Ø
authors.initials: PØ
authors.lastname: Falnes
authors.forename: Jon K
authors.initials: JK
authors.lastname: Laerdahl
authors.forename: Magnar
authors.initials: M
authors.lastname: Bjørås
authors.forename: Ingrun
authors.initials: I
authors.lastname: Alseth
mesh_terms: D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics; D015870:Gene Expression; D006899:Mixed Function Oxygenases / Q000235:genetics; D009154:Mutation; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism*; D021381:Protein Transport; D012568:Schizosaccharomyces / Q000201:enzymology* / Q000235:genetics; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism*; D017422:Sequence Analysis, DNA
chemical_list: D029968:Escherichia coli Proteins; D029702:Schizosaccharomyces pombe Proteins; D006899:Mixed Function Oxygenases; C466427:AlkB protein, E coli; D010089:Oxidoreductases, N-Demethylating; C574547:Abh1 protein, S pombe; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase