DNA unwinding upon strand-displacement binding of a thymine-substituted polyamide to double-stranded DNA.
It was recently found that polyamide nucleic acid (PNA) analogues consisting of thymines attached to an aminoethylglycine backbone bind strongly and sequence-selectively to adenine sequences of oligonucleotides and double-stranded DNA [Nielsen, P. E., Egholm, M., Berg, R. H. & Buchardt, O. (1991) Science 254, 1497-1500]. It was concluded that the binding to double-stranded DNA was accomplished via strand displacement, in which the PNA bound to the Watson-Crick complementary adenine-containing strand, whereas the thymine-containing strand was extruded in a virtually single-stranded conformation. This model may provide a general way in which to obtain sequence-specific recognition of any sequence in double-stranded DNA by Watson-Crick hydrogen-bonding base-pair recognition, and it is thus paramount to rigorously establish this binding mode for synthetic DNA-binding ligands. We now report such results from electron microscopy. Furthermore, we show that binding of PNA to closed circular DNA results in unwinding of the double helix corresponding to approximately one turn of the double helix per 10 base pairs. The DNA.PNA complex, which is formed at low salt concentration (only a small portion of DNA molecules show complex formation at NaCl concentration higher than 40 mM), is exceptionally kinetically stable and cannot be dissociated by increasing salt concentration up to 500 mM.
Cherny
D Y
DY
Institute of Molecular Genetics, Russian Academy of Sciences, Moscow.
Belotserkovskii
B P
BP
Frank-Kamenetskii
M D
MD
Egholm
M
M
Buchardt
O
O
Berg
R H
RH
Nielsen
P E
PE
D004247:DNA / Q000737:chemistry*; D004265:DNA Helicases / Q000737:chemistry; D004270:DNA, Circular / Q000737:chemistry; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D009757:Nylons / Q000737:chemistry*; D009994:Osmolar Concentration; D010957:Plasmids
D004270:DNA, Circular; D009757:Nylons; D004247:DNA; D004265:DNA Helicases