Nucleic Acids Research, 2000, Vol. 28, No. 14 2690-2694
© 2000 Oxford University Press
Telomerase-dependent repeat divergence at the 3' ends of yeast telomeres
Swiss Institute for Experimental Cancer Research (ISREC), CH-1066 Epalinges, Switzerland
Yeast telomeres consist of ~300 nt of degenerate repeats with the consensus sequence G2–3(TG)1–6. We developed a method for the amplification of a genetically marked telomere by PCR, allowing precise length and sequence determination of the G-rich strand including the 3' terminus. We examined wild-type cells, telomerase RNA deficient cells and a strain deleted for YKU70, which encodes for a protein involved in telomere maintenance and DNA double strand break repair. The 3' end of the G-rich strand was found to be at a variable position within the telomeric repeat. No preference for either thymine or guanine as the 3' base was detected. Comparison of telomere sequences from clonal populations revealed that telomeres consist of a centromere-proximal region of stable sequence and a distal region with differing degenerate repeats. In wild-type as well as yku70-
cells, variation in the degenerate telomeric repeats was detected starting 40–100 nt from the 3' end. Sequence divergence was abolished after deletion of the telomerase RNA gene. Thus, this region defines the domain where telomere shortening and telomerase-mediated extension occurs. Since this domain is much larger than the number of nucleotides lost per generation in the absence of telomerase, we propose that telomerase does not extend a given telomere in every cell cycle.
* To whom correspondence should be addressed. Tel: +41 21 692 5912; Fax: +41 21 652 6933; Email: joachim.lingner@isrec.unil.ch
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