Selective recognition of G-qQuadruplex telomeric DNA by a bis(quinacridine) macrocycle.
The interaction of G-quadruplex DNA with the macrocyclic compound BOQ1, which possesses two dibenzophenanthroline (quinacridine) subunits, has been investigated by a variety of methods. The oligonucleotide 5'-A(GGGT(2)A)(3)G(3), which mimics the human telomeric repeat sequence and forms an intramolecular quadruplex, was used as one model system. Equilibrium binding constants measured by biosensor surface plasmon resonance (SPR) methods indicate a high affinity of the macrocycle for the quadruplex conformation (K > 1 x 10(7) M(-)(1)) with two equivalent binding sites. The affinity of BOQ1 for DNA duplexes is at least 1 order of magnitude lower. In addition, the macrocycle is more selective than the monomeric control compound (MOQ2), which is not able to discriminate between the two DNA structures (K(duplex) approximately K(quadruplex) approximately 10(6) M(-)(1)). Strong binding of BOQ1 to G4 DNA sequences was confirmed by fluorometric titrations with a tetraplex-forming oligonucleotide. Competition dialysis experiments with a panel of different DNA structures, from single strands to quadruplexes, clearly established the quadruplex binding specificity of BOQ1. Fluorescence resonance energy transfer (FRET) T(m) experiments with a doubly labeled oligonucleotide also revealed a strong stabilization of the G4 conformation in the presence of BOQ1 (DeltaT(m) = +28 degrees C). This DeltaT(m) value is one of the highest values measured for a G-quadruplex ligand and is significantly higher than observed for the monomer control compounds (DeltaT(m) = +10-12 degrees C). Gel mobility shift assays indicated that the macrocycle efficiently induces the formation of G-tetraplexes. Strong inhibition of telomerase was observed in the submicromolar range (IC(50) = 0.13 microM). These results indicate that macrocycles represent an exciting new development opportunity for targeting DNA quadruplexes.
Teulade-Fichou
Marie-Paule
MP
Laboratoire de Chimie des Interactions Moléculaires, Collège de France, CNRS UPR 285, 11 place Marcelin Berthelot, 75005 Paris, France.
Carrasco
Carolina
C
Guittat
Lionel
L
Bailly
Christian
C
Alberti
Patrizia
P
Mergny
Jean-Louis
JL
David
Arnaud
A
Lehn
Jean-Marie
JM
Wilson
W David
WD
D000166:Acridines / Q000737:chemistry*; D001667:Binding, Competitive; D004247:DNA / Q000737:chemistry*; D003956:Dialysis / Q000379:methods; D031541:Fluorescence Resonance Energy Transfer; D054856:G-Quadruplexes; D007700:Kinetics; D009841:Oligonucleotides / Q000737:chemistry; D010618:Phenanthrolines / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity; D020349:Surface Plasmon Resonance; D019098:Telomerase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D016615:Telomere / Q000737:chemistry* / Q000235:genetics
D000166:Acridines; D009841:Oligonucleotides; D010618:Phenanthrolines; D004247:DNA; D019098:Telomerase