Single-molecule analysis of human telomerase monomer |
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Authors: | Alves David Li Haitao Codrington Rosalind Orte Angel Ren Xiaojun Klenerman David Balasubramanian Shankar |
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Affiliation: | University Chemical Laboratories, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK. |
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Abstract: | Human telomerase is a ribonucleoprotein that is minimally comprised of protein (hTERT) and RNA (hTR) components. We have applied single-molecule fluorescence two-color coincidence detection to characterize complex formation between fluorophore-labeled components in solution. By systematic labeling and in vitro assembly of hTERT, hTR and telomerase's DNA substrate, we have established that catalytically functional human telomerase comprises a stable hTERT:hTR:substrate interaction in a 1:1:1 absolute stoichiometry. |
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