Telomere fusions caused by mutating the terminal region of telomeric DNA

Mutations in the template region of a telomerase RNA gene can lead to the c orresponding sequence alterations appearing in newly synthesized telomeric repeats. We analyzed a set of mutations in the template region of the telom erase RNA gene (TER1) of the budding yeast Kluyveromyces lactis that were p redicted to lead to synthesis of mutant telomeric repeats with disrupted bi nding of the telomeric protein Rap1p. We showed previously that mutating th e left side of the 12-bp consensus Rap1p binding site led to immediate and severe telomere elongation. Here, we show that, in contrast, mutating eithe r the right side of the site or both sides together leads initially to telo mere shortening. On additional passaging, certain mutants of both categorie s exhibit telomere-telomere fusions, Often, six new Bal-31-resistant. telom ere repeat-containing bands appeared, and we infer that each of the six K. lactis chromosomes became circularized. These fusions were not stable, appe aring occasionally to resolve and then reform. We demonstrate directly that a linear minichromosome introduced into one of the fusion mutant strains c ircularized by means of end-to-end fusions of the mutant repeat tracts. In contrast to the chromosomal circularization reported previously in Schizosa ccharomyces pombe mutants defective in telomere maintenance, the K. lactis telomere fusions retained their telomeric DNA repeat sequences.