Intramolecular triplex formation of the purine.purine.pyrimidine type.
Six octadecamers with hairpin motifs have been synthesized and investigated for possible intramolecular triplex formation. Electrophoretic, hypochromic, and CD evidence suggest that d(CCCCTTTGGGGTTTGGGG) and d(GGGGTTTGGGGTTTCCCC) can form G.G.C intramolecular triplexes via double hairpin formation in neutral solutions, presumably with the terminal G tract folding back along the groove of the hairpin duplex. In contrast, d(GGGGTTTCCCCTTTGGGG) and the three corresponding 18-mers containing one G and two C tracts each forms a single hairpin duplex with a dangling single strand. The design of the sequences has led to the conclusion that the two G tracts are antiparallel to each other in such a triplex. Magnesium chloride titrations indicate that Mg2+ is not essential for such an intramolecular triplex formation. The main advantage of our constructs when compared to the intermolecular triplex formation is that the shorter triplex stem can be formed in a much lower DNA concentration. The merit of G.G.C triplex, in contrast to that of C+.G.C, lies in the fact that acidic condition is not required in its formation and will, thus, greatly expand our repertoire in the triplex strategy for the recognition and cleavage of duplex DNA. Spectral binding studies with actinomycin D (ACTD) and chromomycin A3 (CHR) as well as fluorescence lifetime measurements with ethidium bromide (EB) suggest that although hairpin duplexes bind these drugs quite well, the intramolecular triplexes bind poorly. Interestingly, the binding densities for the strong-binding hairpins obtained from Scatchard plots are about one ACTD molecule per oligomeric strand, whereas more than two drug molecules are found in the case of CHR, in agreement with the recent NMR studies indicating that CHR binds to DNA in the form of a dimer.
Chen
F M
FM
Department of Chemistry, Tennessee State University, Nashville 37209-1561.
D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D014128:Chromomycin A3 / Q000494:pharmacology; D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry* / Q000187:drug effects; D003609:Dactinomycin / Q000494:pharmacology; D004996:Ethidium / Q000494:pharmacology; D015636:Magnesium Chloride / Q000494:pharmacology; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation / Q000187:drug effects; D011687:Purines / Q000737:chemistry*; D011740:Pyrimidine Dimers; D011743:Pyrimidines / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
D011687:Purines; D011740:Pyrimidine Dimers; D011743:Pyrimidines; D015636:Magnesium Chloride; D003609:Dactinomycin; D004247:DNA; D014128:Chromomycin A3; D004996:Ethidium