title: New synthetic substrates of mammalian nucleotide excision repair system.
abstract: DNA probes for the studies of damaged strand excision during the nucleotide excision repair (NER) have been designed using the novel non-nucleosidic phosphoramidite reagents that contain N-[6-(9-antracenylcarbamoyl)hexanoyl]-3-amino-1,2-propandiol (nAnt) and N-[6-(5(6)-fluoresceinylcarbamoyl)hexanoyl]-3-amino-1,2-propandiol (nFlu) moieties. New lesion-imitating adducts being inserted into DNA show good substrate properties in NER process. Modified extended linear nFlu- and nAntr-DNA are suitable for estimation of specific excision activity catalysed with mammalian whole-cell extracts. The following substrate activity range was revealed for the model 137-bp linear double-stranded DNA: nAnt-DNA ≈ nFlu-DNA > Chol-DNA (Chol-DNA--legitimate NER substrate that contains non-nucleoside fragment bearing cholesterol residue). In vitro assay shows that modified DNA can be a useful tool to study NER activity in whole-cell extracts. The developed approach should be of general use for the incorporation of NER-sensitive distortions into model DNAs. The new synthetic extended linear DNA containing bulky non-nucleoside modifications will be useful for NER mechanism study and for applications.
authors.affiliation: Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Sciences, Novosibirsk 630090, Russia.
authors.forename: Alexey
authors.initials: A
authors.lastname: Evdokimov
authors.forename: Irina
authors.initials: I
authors.lastname: Petruseva
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Tsidulko
authors.forename: Ludmila
authors.initials: L
authors.lastname: Koroleva
authors.forename: Inna
authors.initials: I
authors.lastname: Serpokrylova
authors.forename: Vladimir
authors.initials: V
authors.lastname: Silnikov
authors.forename: Olga
authors.initials: O
authors.lastname: Lavrik
mesh_terms: D000818:Animals; D000873:Anthracenes / Q000737:chemistry*; D016466:CHO Cells; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D015342:DNA Probes / Q000138:chemical synthesis / Q000737:chemistry*; D004260:DNA Repair; D005452:Fluoresceins / Q000737:chemistry*; D006801:Humans; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry; D010755:Organophosphates / Q000737:chemistry*; D009943:Organophosphorus Compounds / Q000737:chemistry
chemical_list: D000873:Anthracenes; D015342:DNA Probes; D005452:Fluoresceins; D009838:Oligodeoxyribonucleotides; D010755:Organophosphates; D009943:Organophosphorus Compounds; C434331:phosphoramidite; D004247:DNA