New light-sensitive nucleosides for caged DNA strand breaks.
Phototriggered bod cleavage has found wide application in chemistry as well as in biology. Nevertheless, there are only a few methods available for site-specific photochemical induction of DNA strand scission despite numerous potential applications. In this study we report the development of new photocleavable nucleotides based on the photochemistry of o-nitrobenzyl esters. The light-sensitive moieties were generated through introduction of o-nitrophenyl groups at the 5'C position of the nucleoside sugar backbone. The newly synthesized, modified nucleosides were incorporated in oligonucleotides and are able to build stable DNA duplexes. In such a way modified oligonucleotides ca cleaved site-specifically upon irradiation with > 360 nm light with high efficiency. Furthermore, we show that these modifications can be bypassed in DNA synthesis promoted by Thermus aquaticus DNA polymerase.
Dussy
Adrian
A
Kekulé-Institut für Organische Chemie und Biochemie, Universität Bonn, Gerhard-Domagk-Strasse I, 53121 Bonn, Germany.
Meyer
Christoph
C
Quennet
Edith
E
Bickle
Thomas A
TA
Giese
Bernd
B
Marx
Andreas
A
D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence; D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry* / Q000528:radiation effects; D004249:DNA Damage; D017931:DNA Primers; D004261:DNA Replication; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics / Q000528:radiation effects; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D008027:Light; D009705:Nucleosides / Q000528:radiation effects; D013824:Thermus / Q000201:enzymology / Q000235:genetics
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D009705:Nucleosides; D004247:DNA; D004259:DNA-Directed DNA Polymerase