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Originally published as MBC in Press, 10.1091/mbc.E02-06-0327 on October 16, 2002
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Vol. 14, Issue 1, 1-13, January 2003

N-terminal Domain of Yeast Telomerase Reverse Transcriptase: Recruitment of Est3p to the Telomerase Complex

Katherine L. Friedman,*dagger Jeremy J. Heit,Dagger § David M. Long, and Thomas R. CechDagger

 *Vanderbilt University, Department of Biological Sciences, Nashville, Tennessee 37235;  Dagger Department of Chemistry and Biochemistry and Howard Hughes Medical Institute, University of Colorado, Boulder, Colorado 80309-0215; and  Department of Plant Sciences, Montana State University, Bozeman, Montana 59717

Telomerase is a reverse transcriptase that maintains chromosome ends. The N-terminal half of the catalytic protein subunit (TERT) contains three functional domains (I, II, and III) that are conserved among TERTs but not found in other reverse transcriptases. Guided by an amino acid sequence alignment of nine TERT proteins, mutations were introduced into yeast TERT (Est2p). In support of the proposed alignment, mutation of virtually all conserved residues resulted in loss-of-function or temperature sensitivity, accompanied by telomere shortening. Overexpression of telomerase component Est3p led to allele-specific suppression of the temperature-sensitive mutations in region I, suggesting that Est3p interacts with this protein domain. As predicted by the genetic results, a lethal mutation in region I resulted in loss of Est3p from the telomerase complex. We conclude that Est2p region I is required for the recruitment of Est3p to yeast telomerase. Given the phylogenetic conservation of region I of TERT, this protein domain may provide the equivalent function in all telomerases.


Online version of this article contains dataset material. Online version is available at www.molbiolcell.org.

dagger Corresponding author. E-mail address: katherine.friedman{at}vanderbilt.edu.

§ Present address: Department of Developmental Biology, Beckman B300, Stanford, CA 94305-5329.


Molecular Biology of the Cell
Vol. 14, 1-13, January 2003
Copyright © 2003 by The American Society for Cell Biology



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