In search of a Hoogsteen base paired DNA duplex in aqueous solution.
When the oligodeoxynucleotides d(A)6 and d(T)6 are mixed together in a 1:1 ratio (in 100 mM NaCl), the NH signals in the NMR spectrum gave a typical signature of Watson-Crick paired (WC) and Hoogsteen paired (H) AT base pairs. The observation indicates two schemes: Scheme I, WC and H duplexes in slow equilibrium, i.e., WC in equilibrium with H, Scheme II, the WC helix formed is unstable and that it disproportionates into a triple helix (TR) and free d(A)6. We show that (i) addition of extra d(A)6 does not change the helix composition, (ii) addition of a minor-groove specific drug Dst2 (a distamycin analogue) results in an exclusive WC helix-drug duplex, while it does not destabilize triple helix in a 1:2 mixture. In addition we have compared the melting profile, 31P NMR spectra, 1H NMR spectra and the salt dependence of the 1:1 mixture and that of a pure triple helix. All the data from the above experiments overwhelmingly favor Scheme I. However Scheme II cannot be categorically excluded. Based on 1D/2D NMR studies, we have characterized the structural properties of the Hoogsteen double helix in terms of nucleotide conformations. In addition, we computationally demonstrate that the relative stability of the WC over the H duplexes increases with increasing chain length.
Sarma
M H
MH
Department of Chemistry, State University of New York, Albany 12222.
Umemoto
K
K
Gupta
G
G
Luo
J
J
Sarma
R H
RH
D000225:Adenine / Q000737:chemistry; D001482:Base Composition; D004247:DNA / Q000737:chemistry*; D004214:Distamycins; D015636:Magnesium Chloride; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D012965:Sodium Chloride; D012996:Solutions; D013696:Temperature; D013941:Thymine / Q000737:chemistry
D004214:Distamycins; D012996:Solutions; D015636:Magnesium Chloride; D012965:Sodium Chloride; D004247:DNA; D000225:Adenine; D013941:Thymine