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Vol. 12, Issue 12, 4078-4089, December 2001

Repair of Chromosome Ends after Telomere Loss in Saccharomyces

Jeff L. Mangahas, Mary Kate Alexander, Lisa L. Sandell, and Virginia A. Zakian*

Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544

Removal of a telomere from yeast chromosome VII in a strain having two copies of this chromosome often results in its loss. Here we show that there are three pathways that can stabilize this broken chromosome: homologous recombination, nonhomologous end joining, and de novo telomere addition. Both in a wild-type and a recombination deficient rad52 strain, most stabilization events were due to homologous recombination, whereas nonhomologous end joining was exceptionally rare. De novo telomere addition was relatively rare, stabilizing <0.1% of broken chromosomes. Telomere addition took place at a very limited number of sites on chromosome VII, most occurring close to a 35-base pair stretch of telomere-like DNA that is normally ~50 kb from the left telomere of chromosome VII. In the absence of the Pif1p DNA helicase, telomere addition events were much more frequent and were not concentrated near the 35-base pair tract of telomere-like DNA. We propose that internal tracts of telomere-like sequence recruit telomerase by binding its anchor site and that Pif1p inhibits telomerase by dissociating DNA primer-telomerase RNA interactions. These data also show that telomeric DNA is essential for the stable maintenance of linear chromosomes in yeast.

* Corresponding author. E-mail address: vzakian{at}molbio.princeton.edu.

Molecular Biology of the Cell
Vol. 12, 4078-4089, December 2001
Copyright © 2001 by The American Society for Cell Biology


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