Abstract
Several photoactive groups have been covalently attached to the 3’- or 5’-ends of oligodeoxynucleotides. These include proflavine, azidoderivatives (azidophenacyl, azidoproflavine), porphyrins, 9-aminoacridine, ellipticine and diazapyrene derivatives. These derivatized oligonucleotides bind to their complementary sequences on single-stranded nucleic acids. Upon visible or near-UV irradiation of these complexes the oligonucleotide becomes covalently crosslinked to the target nucleic acid. Upon treatment by 1 M piperidine at 90°C the photocrosslinked species are converted to cleaved products. The location of the cleavage sites identifies the nucleotides that were involved in the photocrosslinking reactions. Only ellipticine and diazapyrene derivatives led to cleavage reactions at neutral pH.
Homopyrimidine oligodeoxynucleotides carrying a photoactive group can bind to the major groove of duplex DNA at homopurine.Homopyrimidine sequences. A local triple helix is formed. Upon visible or near-UV irradiation crosslinking of the oligonucleotide is observed on both strands of the double helix. Cleavage reactions occur at the photocrosslinking sites upon alkali treatment. The homopyrimidine oligonucleotide is bound (hydrogen-bonded) in a parallel orientation with respect to the purine-containing strand of double-helical DNA.
Oligodeoxynucleotides can be synthesized with the [ α]-anomers of nucleoside units instead of the natural [β]-anomers. 01igo-[α]-deoxynucleotides are resistant to nucleases and form hybrids with complementary natural ([ β;]) sequences. Photocrosslinking and cleavage reactions were used to demonstrate that the two strands of an [β;]-] (DNA)-[α;](DNA) double helix adopt a parallel orientation. An [β;]-octathymidylate is able to bind to the major groove of duplex DNA containing a d(A<Subscript>8</Subscript>).d(T<Subscript>8</Subscript>) sequence. Its orientation is parallel to the purine-containing strand as is the corresponding [α] analogue.
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© 1989 Kluwer Academic Publishers
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Hélène, C., Doan, T.L., Thuong, N.T. (1989). Sequence-Targeted Photochemical Reactions in Single-Stranded and Double-Stranded Nucleic Acids by Oligonucleotide-Photosensitizer Conjugates. In: Nielsen, P.E. (eds) Photochemical Probes in Biochemistry. NATO ASI Series, vol 272. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-0925-0_17
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DOI: https://doi.org/10.1007/978-94-009-0925-0_17
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