Cell
Volume 166, Issue 5, 25 August 2016, Pages 1188-1197.e9
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Article
Live Cell Imaging Reveals the Dynamics of Telomerase Recruitment to Telomeres

https://doi.org/10.1016/j.cell.2016.07.033Get rights and content
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Highlights

  • Live cell, single-molecule imaging of doubly genome-edited cell lines

  • Telomerase uses three-dimensional diffusion to search for telomeres

  • Telomerase forms short, dynamic and long, static interactions with telomeres

  • Observations provide a new model for human telomerase recruitment to telomeres

Summary

Telomerase maintains genome integrity by adding repetitive DNA sequences to the chromosome ends in actively dividing cells, including 90% of all cancer cells. Recruitment of human telomerase to telomeres occurs during S-phase of the cell cycle, but the molecular mechanism of the process is only partially understood. Here, we use CRISPR genome editing and single-molecule imaging to track telomerase trafficking in nuclei of living human cells. We demonstrate that telomerase uses three-dimensional diffusion to search for telomeres, probing each telomere thousands of times each S-phase but only rarely forming a stable association. Both the transient and stable association events depend on the direct interaction of the telomerase protein TERT with the telomeric protein TPP1. Our results reveal that telomerase recruitment to telomeres is driven by dynamic interactions between the rapidly diffusing telomerase and the chromosome end.

Keywords

CRISPR-Cas9
diffusion constant
dynamics
genome editing
Halo-tag
TERT
TPP1
live cell imaging
telomerase recruitment
telomeres

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