title: Oxidative demethylation of 3-methylthymine and 3-methyluracil in single-stranded DNA and RNA by mouse and human FTO.
abstract: The human obesity susceptibility gene, FTO, encodes a protein that is homologous to the DNA repair AlkB protein. The AlkB family proteins utilize iron(II), alpha-ketoglutarate (alpha-KG) and dioxygen to perform oxidative repair of alkylated nucleobases in DNA and RNA. We demonstrate here the oxidative demethylation of 3-methylthymine (3-meT) in single-stranded DNA (ssDNA) and 3-methyluracil (3-meU) in single-stranded RNA (ssRNA) by recombinant human FTO protein in vitro. Both human and mouse FTO proteins preferentially repair 3-meT in ssDNA over other base lesions tested. They showed negligible activities against 3-meT in double-stranded DNA (dsDNA). In addition, these two proteins can catalyze the demethylation of 3-meU in ssRNA with a slightly higher efficiency over that of 3-meT in ssDNA, suggesting that methylated RNAs are the preferred substrates for FTO.
authors.affiliation: Department of Chemistry, The University of Chicago, 929 East 57th Street, Chicago, IL 60637, USA.
authors.forename: Guifang
authors.initials: G
authors.lastname: Jia
authors.forename: Cai-Guang
authors.initials: CG
authors.lastname: Yang
authors.forename: Shangdong
authors.initials: S
authors.lastname: Yang
authors.forename: Xing
authors.initials: X
authors.lastname: Jian
authors.forename: Chengqi
authors.initials: C
authors.lastname: Yi
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Zhou
authors.forename: Chuan
authors.initials: C
authors.lastname: He
mesh_terms: D000071516:Alpha-Ketoglutarate-Dependent Dioxygenase FTO; D000818:Animals; D004260:DNA Repair; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008745:Methylation; D051379:Mice; D006899:Mixed Function Oxygenases; D010084:Oxidation-Reduction; D007652:Oxo-Acid-Lyases / Q000737:chemistry / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry; D013379:Substrate Specificity; D013941:Thymine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D014498:Uracil / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism
chemical_list: D004277:DNA, Single-Stranded; D011506:Proteins; D011994:Recombinant Proteins; D014498:Uracil; C055991:3-methyluracil; D012313:RNA; D006899:Mixed Function Oxygenases; C524916:FTO protein, mouse; D000071516:Alpha-Ketoglutarate-Dependent Dioxygenase FTO; C526492:FTO protein, human; D007652:Oxo-Acid-Lyases; D013941:Thymine; C002347:3-methylthymine
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