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. 2011 May 17;39(16):7161–7178. doi: 10.1093/nar/gkr234

Figure 6.

Figure 6.

G4R1/RHAU-catalyzed resolution of 5′-TAMRA-labeled tetramolecular G4-DNA substrate is inhibited most efficiently by unlabeled unimolecular c-Myc 51-mer G4-DNA. (A, C and E) Phosphorimage of representative (of three repetitions) nondenaturing gel electropherograms; the top triangles indicate increasing concentrations of unlabeled (A) Poly A Zic1 47-mer G4-DNA, (C) c-Myc 51-mer G4-DNA, or (E) tetramolecular Z33 G4-DNA added to G4R1/RHAU-catalyzed resolving reactions of 5′-TAMRA-labeled tetramolecular Z33 G4-DNA substrate. Lane 1 of panels A, C and E: 2 nM 5′-TAMRA-labeled tetramolecular Z33 G4-DNA substrate in the absence of enzyme or unlabeled unimolecular G4-DNA. Lanes 2–12 of panels A, C and E: 2 nM 5′-TAMRA-labeled tetramolecular Z33 G4-DNA substrate after incubation with 1 unit of G4R1/RHAU plus increasing concentrations of unlabeled inhibitor G4-DNA. (B, D and F) Graphic representation of concentration-dependent inhibition of G4R1/RHAU-catalyzed resolution of 5′-TAMRA-labeled tetramolecular Z33 G4-DNA substrate by unlabeled (B) Poly A Zic1 47-mer G4-DNA, (D) c-Myc 51-mer G4-DNA, or (F) tetramolecular Z33 G4-DNA. Inhibition is represented by the percentage of maximal resolution (error bars are Mean ± SD, n = 3 independent experiments). Inhibition of 50% is observed at a 6.25:1 unimolecular Poly A Zic1 DNA 47-mer G4-DNA:Z33 tetramer G4-DNA molar ratio, at a 0.06:1 unimolecular c-Myc DNA 51-mer:Z33 tetramer G4-DNA molar ratio, and at a 1:1 unlabeled Z33 tetramer G4-DNA:labeled Z33 tetramer G4-DNA molar ratio.