RESUMO
A deeper investigation of the chemistry that occurs on the newly discovered epigenetic DNA bases 5-hydroxymethyl-(hmdC), 5-formyl-(fdC), and 5-carboxy-deoxycytidine (cadC) requires chemical tool compounds, which are able to dissect the different potential reaction pathways in cells. Here we report that the 2'-(R)-fluorinated derivatives F-hmdC, F-fdC, and F-cadC, which are resistant to removal by base excision repair, are good substrates for DNA polymerases and TET enzymes. This result shows that the fluorinated compounds are ideal tool substances to investigate potential C-C-bond cleaving reactions in the context of active demethylation.
Assuntos
Citidina/análogos & derivados , Citidina/metabolismo , DNA Polimerase Dirigida por DNA/metabolismo , DNA/metabolismo , Dioxigenases/metabolismo , Polifosfatos/metabolismo , Citidina/genética , DNA/química , DNA/genética , Epigênese Genética , Células HEK293 , Halogenação , Humanos , Polifosfatos/química , Especificidade por SubstratoRESUMO
Real-time enzymatic studies are gaining importance as their chemical and technical instrumentation improves. Here we report the efficient synthesis of γ-alkyne modified triphosphate amidates that are converted into a variety of γ-fluorophore labeled triphosphates by Cu(I) catalyzed alkyne/azide click reactions. The synthesized triphosphates are incorporated into DNA by DNA polymerases.
Assuntos
Química Click , Nucleotídeos/química , Polifosfatos/química , Polifosfatos/síntese química , Alcinos/química , Azidas/química , Sequência de Bases , Catálise , Cobre/química , DNA/síntese química , DNA/química , DNA/genéticaRESUMO
The development of metal base pairs is of immense importance for the construction of DNA nanostructures. Here we report the synthesis of a biaryl pyrazole-phenol nucleoside that forms in DNA a stable self-pair upon complexation of a Cu(II) ion. A sequence with five consecutive pyrazole nucleotides allows the complexation of five Cu(II) ions in a row.
