title: Incorporation of non-nucleoside triphosphate analogues opposite to an abasic site by human DNA polymerases beta and lambda.
abstract: A novel class of non-nucleoside triphosphate analogues, bearing hydrophobic groups sterically similar to nucleosides linked to the alpha-phosphate but lacking the chemical functional groups of nucleic acids, were tested against six different DNA polymerases (polymerases). Human polymerases alpha, beta and lambda, and Saccharomyces cerevisiae polymerase IV, were inhibited with different potencies by these analogues. On the contrary, Escherichia coli polymerase I and HIV-1 reverse transcriptase were not. Polymerase beta incorporated these derivatives in a strictly Mn++-dependent manner. On the other hand, polymerase lambda could incorporate some alkyltriphosphate derivatives with both Mg++ and Mn++, but only opposite to an abasic site on the template strand. The active site mutant polymerase lambda Y505A showed an increased ability to incorporate the analogues. These results show for the first time that neither the base nor the sugar moieties of nucleotides are required for incorporation by family X DNA polymerases.
authors.affiliation: Istituto di Genetica Molecolare IGM-CNR via Abbiategrasso 207, I-27100 Pavia, Italy.
authors.forename: Emmanuele
authors.initials: E
authors.lastname: Crespan
authors.forename: Samantha
authors.initials: S
authors.lastname: Zanoli
authors.forename: Anastasiya
authors.initials: A
authors.lastname: Khandazhinskaya
authors.forename: Igor
authors.initials: I
authors.lastname: Shevelev
authors.forename: Maxim
authors.initials: M
authors.lastname: Jasko
authors.forename: Ludmila
authors.initials: L
authors.lastname: Alexandrova
authors.forename: Marina
authors.initials: M
authors.lastname: Kukhanova
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Blanca
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Villani
authors.forename: Ulrich
authors.initials: U
authors.lastname: Hübscher
authors.forename: Silvio
authors.initials: S
authors.lastname: Spadari
authors.forename: Giovanni
authors.initials: G
authors.lastname: Maga
mesh_terms: D004247:DNA / Q000096:biosynthesis; D004253:DNA Nucleotidylexotransferase / Q000378:metabolism; D004256:DNA Polymerase I / Q000378:metabolism; D019951:DNA Polymerase beta / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry*; D006801:Humans; D008345:Manganese / Q000737:chemistry; D019384:Nucleic Acid Synthesis Inhibitors; D009711:Nucleotides / Q000378:metabolism; D017354:Point Mutation; D011122:Polyphosphates / Q000737:chemistry* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D013698:Templates, Genetic
chemical_list: D004791:Enzyme Inhibitors; D019384:Nucleic Acid Synthesis Inhibitors; D009711:Nucleotides; D011122:Polyphosphates; D029701:Saccharomyces cerevisiae Proteins; D008345:Manganese; D004247:DNA; C415367:DNA polymerase beta2; D004253:DNA Nucleotidylexotransferase; D004256:DNA Polymerase I; D019951:DNA Polymerase beta; D004259:DNA-Directed DNA Polymerase; C084892:POL4 protein, S cerevisiae; C005692:triphosphoric acid