Single-molecule real-time detection of telomerase extension activity

Hwang, Helen; Opresko, Patricia; Myong, Sua

doi:10.1038/srep06391

i

Single-molecule real-time detection of telomerase extension activity

Supporting Files

Sep 29 2014
By Hwang, Helen ; Opresko, Patricia ; Myong, Sua

File Language:

English

Details

Alternative Title:

Sci Rep
Personal Author:

Hwang, Helen ; Opresko, Patricia ; Myong, Sua
Description:

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.
Subjects:

Aminopeptidases Animals Chromosomes Dipeptidyl-Peptidases And Tripeptidyl-Peptidases DNA Replication HEK293 Cells Humans Neoplasms Serine Proteases Telomerase Telomere Telomere-Binding Proteins Telomere Homeostasis
Source:

Sci Rep. 2014; 4.
Pubmed ID:

25263700
Pubmed Central ID:

PMC4178293
Document Type:

Journal Article
Funding:

1F30CA174323-01/CA/NCI NIH HHS/United States ; 343 NIH 1 DP2 GM105453/DP/NCCDPHP CDC HHS/United States ; F30 CA174323/CA/NCI NIH HHS/United States ; R01 ES022944/ES/NIEHS NIH HHS/United States ; R01ES022944/ES/NIEHS NIH HHS/United States
Volume:

4
Collection(s):

CDC Public Access
Main Document Checksum:

urn:sha256:a228adfbfdf59f822e956bd9aa1c5b033da98f67f8fa255a2256468241764c1d
Download URL:

https://stacks.cdc.gov/view/cdc/32196/cdc_32196_DS1.pdf
File Type:

[PDF - 1.27 MB ]

File Language:

English

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