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Structural heterogeneity in intramolecular DNA triple helices

Item Type:Article
Title:Structural heterogeneity in intramolecular DNA triple helices
Creators Name:Weisz, K., Leitner, D., Krafft, C. and Welfle, H.
Abstract:Oligodeoxynucleotides designed to form intramolecular triple helices are widely used as model systems in thermodynamic and structural studies. We now report results from UV, Raman and NMR experiments demonstrating that the strand polarity, which also determines the orientation of the connecting loops, has a considerable impact on the formation and stability of pyr·pur·pyr triple helices. There are two types of monomolecular triplexes that can be defined by the location of their purine tract at either the 5'-or 3'-end of the sequence. We have examined four pairs of oligonucleotides with the same base composition but with reserved polarity that can fold into intramolecular triple helices with seven base triplets and two T4 loops under appropriate conditions. UV spectroscopic monitoring of thermal denaturation indicates a consistently higher thermal stability for the 5'-sequences at pH 5.0 in the absence of Mg2+ ions. Raman spectra provide evidence for the formation of triple helices at pH 5 for oligomers with purine tracts located at either the 5'- or 3'-end of the sequence. However, NMR measurements reveal considerable differences in the secondary structures formed by the two types of oligonucleotides. Thus, at acidic pH significant structural heterogeneity is observed for the 3'-sequences. Employing selectively 15N-labeled oligomers, NMR experiments indicate a folding pattern for the competing structures that at least partially changes both Hoogsteen and Watson-Crick base-base interactions.
Keywords:DNA Loop, NMR, Raman Spectroscopy, Strand Polarity, UV Melting
Source:Biological Chemistry
ISSN:1431-6730
Publisher:de Gruyter
Volume:381
Number:4
Page Range:275-283
Date:1 April 2000
Official Publication:https://doi.org/10.1515/BC.2000.036
PubMed:View item in PubMed

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