Nucleic Acids Research, Vol 24, Issue 16 3181-3188, Copyright © 1996 by Oxford University Press
B Faucon, JL Mergny and C Helene
Exon 5 of the human aprt gene contains an oligo-purine-oligopyrimidine
stretch of 17 bp (5'-CCCTCTTCTCTCTCCT-3') within the coding region. (T,C)-,
(G,T)- and (G,A)-containing oligonucleotides were compared for their
ability to form stable triple helices with their DNA target. (G,T)
oligodeoxynucleotides, whether parallel or antiparallel, were unable to
bind to this sequence. This is in contrast to (G,A) (purine) and (T,C)
(pyrimidine) oligonucleotides, which bind to the duplex at near neutral pH.
Binding was highly sequence specific, as unrelated competitors were unable
to interfere with target recognition. A major difference between the purine
and pyrimidine oligodeoxynucleotides was observed in the kinetics of
binding: the (G,A) oligonucleotide binds to its target much faster than the
(T,C) oligomer. With the purine oligonucleotide, complete binding was
achieved in a matter of minutes at micromolar concentrations, whereas
several hours were required with the pyrimidine oligomer. Thus, the general
observation that triplex formation is slow with pyrimidine
oligodeoxynucleotides does not hold for (G,A) oligodeoxynucleotides. Purine
and pyrimidine oligodeoxynucleotides covalently linked to a psoralen group
were able to induce crosslinks on the double-stranded DNA target upon UV
irradiation. This study provides a detailed comparison of the different
types of DNA triplexes under the same experimental conditions.
ARTICLES
Effect of third strand composition on the triple helix formation: purine versus pyrimidine oligodeoxynucleotides
Laboratoire de Biophysique, Museum National d'Histoire Naturelle, INSERM U201, CNRS UA481, Paris, France.
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