Single-molecule real-time detection of telomerase extension activity.

Abstract:

:The ends of eukaryotic chromosomes are capped by telomeres which consist of tandem G-rich DNA repeats stabilized by the shelterin protein complex. Telomeres shorten progressively in most normal cells due to the end replication problem. In more than 85% of cancers however, the telomere length is maintained by telomerase; a reverse transcriptase that adds telomeric TTAGGG repeats using its integral RNA template. The strong association between telomerase activity and malignancy in many cancers suggests that telomerase activity could serve as a diagnostic marker. We demonstrate single-molecule, real-time telomerase extension activity observed digitally as the telomeric repeats are added to a substrate. The human telomerase complex pulled down from mammalian cells displays extension activity dependent on dNTP concentration. In complex with the processivity factor, POT1-TPP1, telomerase adds repeats at an accelerated rate and yields longer products. Our assay provides a unique detection platform that enables the study of telomerase kinetics with single molecule resolution.

journal_name

Sci Rep

journal_title

Scientific reports

authors

Hwang H,Opresko P,Myong S

doi

10.1038/srep06391

subject

Has Abstract

pub_date

2014-09-29 00:00:00

pages

6391

issn

2045-2322

pii

srep06391

journal_volume

4

pub_type

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