Dynamics of telomeric DNA turnover in yeast.
Telomerase adds telomeric DNA repeats to telomeric termini using a sequence within its RNA subunit as a template. We characterized two mutations in the Kluyveromyces lactis telomerase RNA gene (TER1) template. Each initially produced normally regulated telomeres. One mutation, ter1-AA, had a cryptic defect in length regulation that was apparent only if the mutant gene was transformed into a TER1 deletion strain to permit extensive replacement of basal wild-type repeats with mutant repeats. This mutant differs from previously studied delayed elongation mutants in a number of properties. The second mutation, TER1-Bcl, which generates a BclI restriction site in newly synthesized telomeric repeats, was indistinguishable from wild type in all phenotypes assayed: cell growth, telomere length, and in vivo telomerase fidelity. TER1-Bcl cells demonstrated that the outer halves of the telomeric repeat tracts turn over within a few hundred cell divisions, while the innermost few repeats typically resisted turnover for at least 3000 cell divisions. Similarly deep but incomplete turnover was also observed in two other TER1 template mutants with highly elongated telomeres. These results indicate that most DNA turnover in functionally normal telomeres is due to gradual replicative sequence loss and additions by telomerase but that there are other processes that also contribute to turnover.
McEachern
Michael J
MJ
Department of Genetics, Life Sciences Building, University of Georgia, Athens, Georgia 30602-7223, USA. mjm@arches.uga.edu
Underwood
Dana Hager
DH
Blackburn
Elizabeth H
EH
D004271:DNA, Fungal / Q000378:metabolism*; D007716:Kluyveromyces / Q000201:enzymology / Q000235:genetics*; D009154:Mutation; D012313:RNA / Q000235:genetics*; D019098:Telomerase / Q000235:genetics*; D016615:Telomere / Q000235:genetics*
D004271:DNA, Fungal; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase