The Rad51 Pathway of Telomerase-Independent Maintenance of Telomeres Can Amplify TG_1-3 Sequences in yku and cdc13 Mutants of Saccharomyces cerevisiae

Abstract

In the yeast Saccharomyces cerevisiae, Cdc13, Yku, and telomerase define three parallel pathways for telomere end protection that prevent chromosome instability and death by senescence. We report here that cdc13-1 yku70Δ mutants generated telomere deprotection-resistant cells that, in contrast with telomerase-negative senescent cells, did not display classical crisis events. cdc13-1 yku70Δ cells survived telomere deprotection by exclusively amplifying TG_1-3 repeats (type II recombination). In a background lacking telomerase (tlc1Δ), this process predominated over type I recombination (amplification of subtelomeric Y′ sequences). Strikingly, inactivation of the Rad50/Rad59 pathway (which is normally required for type II recombination) in cdc13-1 yku70Δ or yku70Δ tlc1Δ mutants, but also in cdc13-1 YKU70⁺ tlc1Δ mutants, still permitted type II recombination, but this process was now entirely dependent on the Rad51 pathway. In addition, delayed senescence was observed in cdc13-1 yku70Δ rad51Δ and cdc13-1 tlc1Δ rad51Δ cells. These results demonstrate that in wild-type cells, masking by Cdc13 and Yku prevents the Rad51 pathway from amplifying telomeric TG_1-3 sequences. They also suggest that Rad51 is more efficient than Rad50 in amplifying the sequences left uncovered by the absence of Cdc13 or Yku70.

ACKNOWLEDGMENTS

We thank Thomas Petes, James Haber, Dan Gottschling, Leland Hartwell, and Maria-Pia Longhese for the gifts of strains and plasmids as well as Matthias Rose and his colleagues at Euroscarf for efficiently conveying strains.

This work was supported by grants from the Association pour la Recherche contre le Cancer and the Comité Départemental de la Savoie de la Ligue Nationale contre le Cancer.