Single-molecule analysis of human telomerase monomer.
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.
Alves
David
D
University Chemical Laboratories, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.
Li
Haitao
H
Codrington
Rosalind
R
Orte
Angel
A
Ren
Xiaojun
X
Klenerman
David
D
Balasubramanian
Shankar
S
D002384:Catalysis; D005453:Fluorescence; D006801:Humans; D013379:Substrate Specificity; D019098:Telomerase / Q000737:chemistry* / Q000378:metabolism
C509186:TERT protein, human; D019098:Telomerase