Figure 3: DNA conformational changes induced by completion of telomere DNA repeat synthesis.
From: Coordinated DNA dynamics during the human telomerase catalytic cycle
(a) FRET distributions constructed with doubly labelled telomerase enzyme harbouring hTR U29-Cy5 (P1 stem) and U42-Cy3 dyes initially bound to 5′ biotinylated 18GGG primer. FRET histograms at each indicated step of the repeat synthesis reaction are shown and fit with single Gaussian functions (red lines) centred at FRET ~0.6 (dotted line). Initial 3′ DNA sequence is highlighted in green and nascent DNA sequence is highlighted in white. (b) FRET distributions constructed from U42-Cy3-labelled telomerase enzyme initially bound to 18GGG/T13-Cy5 DNA primer and extended to each indicated step of the repeat synthesis reaction are shown. Data were fit with either one or two Gaussian functions (red lines) to determine the indicated centres of each FRET distribution. (c) FRET distributions constructed from U42-Cy3-labelled telomerase enzyme harbouring the template mutations UA47-48AU, initially bound to the 18GGG/T13-Cy5 primer and extended to each indicated step of the repeat synthesis reaction are shown. Data were fit with either one or two Gaussian functions (red lines) to determine the indicated centres of each FRET distribution. All histograms in the figure are constructed from at least 500 individual molecules.
