Fig. 4: Single molecule analysis reveals a non-productive telomerase complex after 6-thio-dG addition. | Nature Communications

Fig. 4: Single molecule analysis reveals a non-productive telomerase complex after 6-thio-dG addition.

From: Therapeutic 6-thio-deoxyguanosine inhibits telomere elongation in cancer cells by inducing a non-productive stalled telomerase complex

Fig. 4

a Schematic of the experimental setup. LD555-labeled telomerase is pulled down on Cy5-labeled (TTAGGG)3 substrate and immobilized on a quartz slide via a 5’ biotin. The close proximity of the dyes permits FRET. Telomerase-catalyzed addition of dNTPs to the telomere substrate increases the distance between the dyes, causing a decrease in FRET dependent on the number of repeats added. Created in BioRender. Sanford, S. (2025) https://BioRender.com/wtax6ef. b FRET histograms of complexes prior to telomerase activity (top “stalled”) and following a 30 min ‘mock’ activity incubation lacking dNTPs. c FRET histograms of complexes incubated in the presence of dATP, dTTP, and dGTP collected prior to (top) and 15 and 30 min following dNTP addition. Shift from high to low FRET states reports on telomerase movement away from the acceptor dye during telomere elongation. d FRET histograms of complexes collected prior to (top) and 15 and 30 minutes after dATP, dTTP and 6-thio-dGTP (6dGTP) addition. e FRET histograms of complexes collected prior to (top) and 15 and 30 min after dATP, dTTP, dGTP and a 10-fold excess of 6-thio-dGTP addition. Source data are provided as a Source Data file.

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