title: The hyperthermophilic euryarchaeon Archaeoglobus fulgidus repairs uracil by single-nucleotide replacement.
abstract: Hydrolytic deamination of cytosine to uracil in cellular DNA is a major source of C-to-T transition mutations if uracil is not repaired by the DNA base excision repair (BER) pathway. Since deamination increases rapidly with temperature, hyperthermophiles, in particular, are expected to succumb to such damage. There has been only one report of crenarchaeotic BER showing strong similarities to that in most eukaryotes and bacteria for hyperthermophilic Archaea. Here we report a different type of BER performed by extract prepared from cells of the euryarchaeon Archaeoglobus fulgidus. Although immunodepletion showed that the monofunctional family 4 type of uracil-DNA glycosylase (UDG) is the principal and probably only UDG in this organism, a β-elimination mechanism rather than a hydrolytic mechanism is employed for incision of the abasic site following uracil removal. The resulting 3' remnant is removed by efficient 3'-phosphodiesterase activity followed by single-nucleotide insertion and ligation. The finding that repair product formation is stimulated similarly by ATP and ADP in vitro raises the question of whether ADP is more important in vivo because of its higher heat stability.
authors.affiliation: Faculty of Science and Technology, Department of Mathematics and Natural Sciences, University of Stavanger, N-4036 Stavanger, Norway.
authors.forename: Ingeborg
authors.initials: I
authors.lastname: Knævelsrud
authors.forename: Marivi N
authors.initials: MN
authors.lastname: Moen
authors.forename: Kristin
authors.initials: K
authors.lastname: Grøsvik
authors.forename: Gyri T
authors.initials: GT
authors.lastname: Haugland
authors.forename: Nils-Kåre
authors.initials: NK
authors.lastname: Birkeland
authors.forename: Arne
authors.initials: A
authors.lastname: Klungland
authors.forename: Ingar
authors.initials: I
authors.lastname: Leiros
authors.forename: Svein
authors.initials: S
authors.lastname: Bjelland
mesh_terms: D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism; D019611:Archaeoglobus fulgidus / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004249:DNA Damage; D004260:DNA Repair / Q000502:physiology*; D019641:DNA, Archaeal / Q000235:genetics*; D019848:Gene Expression Regulation, Archaeal / Q000502:physiology; D006358:Hot Temperature; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D014498:Uracil; D051981:Uracil-DNA Glycosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D019843:Archaeal Proteins; D019641:DNA, Archaeal; D014498:Uracil; D051981:Uracil-DNA Glycosidase
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