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Coordinated DNA dynamics during the human telomerase catalytic cycle

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  • Joseph W. Parks

    (University of California)

  • Michael D. Stone

    (University of California
    Center for Molecular Biology of RNA, University of California)

Abstract

The human telomerase reverse transcriptase (hTERT) utilizes a template within the integral RNA subunit (hTR) to direct extension of telomeres. Telomerase exhibits repeat addition processivity (RAP) and must therefore translocate the nascent DNA product into a new RNA:DNA hybrid register to prime each round of telomere repeat synthesis. Here, we use single-molecule FRET and nuclease protection assays to monitor telomere DNA structure and dynamics during the telomerase catalytic cycle. DNA translocation during RAP proceeds through a previously uncharacterized kinetic substep during which the 3′-end of the DNA substrate base pairs downstream within the hTR template. The rate constant for DNA primer realignment reveals this step is not rate limiting for RAP, suggesting a second slow conformational change repositions the RNA:DNA hybrid into the telomerase active site and drives the extrusion of the 5′-end of the DNA primer out of the enzyme complex.

Suggested Citation

  • Joseph W. Parks & Michael D. Stone, 2014. "Coordinated DNA dynamics during the human telomerase catalytic cycle," Nature Communications, Nature, vol. 5(1), pages 1-10, September.
    • Handle: RePEc:nat:natcom:v:5:y:2014:i:1:d:10.1038_ncomms5146
    DOI: 10.1038/ncomms5146
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