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Cell. 2016 Aug 25;166(5):1188-1197.e9. doi: 10.1016/j.cell.2016.07.033. Epub 2016 Aug 11.

Live Cell Imaging Reveals the Dynamics of Telomerase Recruitment to Telomeres.

Author information

1
Howard Hughes Medical Institute, Department of Chemistry and Biochemistry, BioFrontiers Institute, University of Colorado Boulder, Boulder, CO 80309, USA.
2
Howard Hughes Medical Institute, Department of Chemistry and Biochemistry, BioFrontiers Institute, University of Colorado Boulder, Boulder, CO 80309, USA. Electronic address: thomas.cech@colorado.edu.

Abstract

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

PMID:
27523609
PMCID:
PMC5743434
DOI:
10.1016/j.cell.2016.07.033
[Indexed for MEDLINE]
Free PMC Article

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