title: New chemically reactive dsDNAs containing single internucleotide monophosphoryldithio links: reactivity of 5'-mercapto-oligodeoxyribonucleotides.
abstract: Novel modified DNA duplexes with single bridging 5'-SS-monophosphoryldithio links [-OP(=O)-O(-)-SS-CH(2)-] were synthesized by autoligation of an oligonucleotide 3'-phosphorothioate and a 5'-mercapto-oligonucleotide previously converted to a 2-pyridyldisulfide adduct. Monophosphoryldisulfide link formation is not a stringent template-dependent process under the conditions used and does not require strong binding of the reactive oligomers to the complementary strand. The modified internucleotide linkage, resembling the natural phosphodiester bond in size and charge density, is stable in water, easily undergoes thiol-disulfide exchange and can be specifically cleaved by the action of reducing reagents. DNA molecules containing an internal -OP(=O)-O(-)-SS-CH(2)- bridge are stable to spontaneous exchange of disulfide-linked fragments (recombination) even in the single-stranded state and are promising reagents for autocrosslinking with cysteine-containing proteins. The chemical and supramolecular properties of oligonucleotides with 5'-sulfhydryl groups were further characterized. We have shown that under the conditions of chemical ligation the 5'-SH group of the oligonucleotide has a higher reactivity towards N-hydroxybenzotriazole-activated phosphate in an adjacent oligonucleotide than does the OH group. This autoligation, unlike disulfide bond formation, proceeds only in the presence of template oligonucleotide, necessary to provide the activated phosphate in close proximity to the SH-, OH- or phosphate function.
authors.affiliation: Department of Chemistry, Lomonosov Moscow State University, Moscow 119899, Russia.
authors.forename: V G
authors.initials: VG
authors.lastname: Metelev
authors.forename: O A
authors.initials: OA
authors.lastname: Borisova
authors.forename: E M
authors.initials: EM
authors.lastname: Volkov
authors.forename: T S
authors.initials: TS
authors.lastname: Oretskaya
authors.forename: N G
authors.initials: NG
authors.lastname: Dolinnaya
mesh_terms: D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D004247:DNA / Q000138:chemical synthesis / Q000737:chemistry*; D004220:Disulfides / Q000737:chemistry; D004229:Dithiothreitol / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000737:chemistry; D008956:Models, Chemical; D009691:Nucleic Acid Denaturation; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D013698:Templates, Genetic; D013873:Thionucleotides / Q000737:chemistry*
chemical_list: D003432:Cross-Linking Reagents; D004220:Disulfides; D009838:Oligodeoxyribonucleotides; D013873:Thionucleotides; D004247:DNA; D005022:Ethyldimethylaminopropyl Carbodiimide; D004229:Dithiothreitol