Real-time detection of human telomerase DNA synthesis by multiplexed single-molecule FRET.

ABSTRACT: Genomic stability in proliferating cells critically depends on telomere maintenance by telomerase reverse transcriptase. Here we report the development and proof-of-concept results of a single-molecule approach to monitor the catalytic activity of human telomerase in real time and with single-nucleotide resolution. Using zero-mode waveguides and multicolor FRET, we recorded the processive addition of multiple telomeric repeats to individual DNA primers. Unlike existing biophysical and biochemical tools, the novel approach enables the quantification of nucleotide-binding kinetics before nucleotide incorporation. Moreover, it provides a means to dissect the unique translocation dynamics that telomerase must undergo after synthesis of each hexameric DNA repeat. We observed an unexpectedly pro

SUBMITTER: Hentschel J 

PROVIDER: S-EPMC10502476 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature