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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Change history
01 May 2008
In the version of this article initially published, parentheses were missing around covalently attached domains in fusion proteins. Also, in the first two lines of the left column of page 288, "hTERT-YFP" should have read "YFP-hTERT" for consistency with the rest of the manuscript. The errors have been corrected in the HTML and PDF versions of the article.
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Acknowledgements
We thank the Leverhulme Trust for supporting this project, the Biotechnology and Biological Sciences Research Council for a PhD studentship (D.A.), and Junta de Andalucia and the European Union 6th Framework for postdoctoral fellowships (A.O.). We thank J. Yeoman for critically proofreading this manuscript.
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D.A., H.L., A.O., S.B. and D.K. conceived and designed the experiments. D.A., H.L., R.C., A.O. and X.R. carried out the experiments. D.A., H.L. and A.O. analyzed the experimental data. All authors discussed the results, and the manuscript was written by D.A. and S.B. with contributions from D.K., H.L. and A.O.
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Supplementary Figures 1–6, Supplementary Tables 1 and 2, Supplementary Methods and Supplementary Discussion (PDF 218 kb)
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Alves, D., Li, H., Codrington, R. et al. Single-molecule analysis of human telomerase monomer. Nat Chem Biol 4, 287–289 (2008). https://doi.org/10.1038/nchembio.82
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DOI: https://doi.org/10.1038/nchembio.82
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