Nucleic Acids Research, Vol 26, Issue 3 787-795, Copyright © 1998 by Oxford University Press
C Autexier and CW Greider
Telomere-specific repeat sequences are essential for chromosome end
stability. Telomerase maintains telomere length by adding sequences de novo
onto chromosome ends. The template domain of the telomerase RNA component
dictates synthesis of species-specific telomeric repeats and other regions
of the RNA have been suggested to be important for enzyme structure and/or
catalysis. Using enzyme reconstituted in vitro with RNAs containing
deletions or substitutions we identified nucleotides in the RNA component
that are important for telomerase activity. Although many changes to
conserved features in the RNA secondary structure did not abolish enzyme
activity, levels of activity were often greatly reduced, suggesting that
regions other than the template play a role in telomerase function. The
template boundary was only altered by changes in stem II that affected the
conserved region upstream of the template, not by changes in other regions,
such as stems I, III and IV, consistent with a role of the conserved region
in defining the 5' boundary of the template. Surprisingly, telomerase RNAs
with substitutions or deletion of residues potentially abolishing the
conserved pseudoknot structure had wild-type levels of telomerase activity.
This suggests that this base pairing interaction may not be required for
telomerase activity per se but may be conserved as a regulatory site for
the enzyme in vivo.
ARTICLES
Mutational analysis of the Tetrahymena telomerase RNA: identification of residues affecting telomerase activity in vitro
Cold Spring Harbor Laboratory, PO Box 100, Cold Spring Harbor, NY 11724, USA.
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