Affordable Access

deepdyve-link
Publisher Website

A single-molecule assay for telomerase structure-function analysis.

Authors
Type
Published Article
Journal
Nucleic Acids Research
Publisher
Oxford University Press
Volume
38
Issue
3
Pages
16–16
Identifiers
DOI: 10.1093/nar/gkp1033
Source
UCSC Cancer biomedical-ucsc
License
Unknown

Abstract

The activity of the telomerase ribonucleoprotein enzyme is essential for the maintenance of genome stability and normal cell development. Despite the biomedical importance of telomerase activity, detailed structural models for the enzyme remain to be established. Here we report a single-molecule assay for direct structural analysis of catalytically active telomerase enzymes. In this assay, oligonucleotide hybridization was used to probe the primer-extension activity of individual telomerase enzymes with single nucleotide sensitivity, allowing precise discrimination between inactive, active and processive enzyme binding events. FRET signals from enzyme molecules during the active and processive binding events were then used to determine the global organization of telomerase RNA within catalytically active holoenzymes. Using this assay, we have identified an active conformation of telomerase among a heterogeneous population of enzymes with distinct structures.

Report this publication

Statistics

Seen <100 times