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. 2023 Dec 28;52(4):1763–1778. doi: 10.1093/nar/gkad1209

Figure 5.

Figure 5.

BG4 addition promotes GQ folding of telomeric constructs containing a destabilizing G substitution. Schematics for biotinylated FRET constructs TELO4 (A), G2T (B), or 8oxoG2 (C) tethered to neutravidin coated slides, followed by 150 nM BG4 addition and then buffer washes. Construct sequences shown in Supplementary Table 2. Histograms of FRET efficiency frequencies observed TELO4 (D), G2T (E), and 8oxoG2 (F) before or after BG4 addition, and sequential buffer wash and SDS addition as indicated. Representative single-molecule traces show FRET efficiencies captured over 90 s. (G) Schematic of SiMPull assay for FRET non-biotinylated FRET TELO4 construct binding to BG4 attached to the slide via FLAG antibody. (H) Histogram of FRET efficiency after adding non-biotinylated TELO4 FRET construct. (I, J) Dwell times for the high-FRET state of G2T and 8oxoG2 constructs in the presence and absence of BG4 from panels E and F. Colors indicate conformations for a range of FRET efficiencies: orange for partially unfolded GQ (marked with U) and blue for folded GQ (marked with F).