Screening for telomeric recombination in wild-typeKluyveromyces lactis

Abstract

Both subtelomeric and telomeric recombination events can be greatly enhanced in Kluyveromyces lactis mutants lacking telomerase and having abnormally short telomeres. In this study, we utilized cells containing a single telomere composed of mutant repeats carrying a phenotypically silent mutation to test whether the exchange of telomeric repeats was a frequent event in mitotic and meiotic wild-type K. lactis cells. Amongst more than 100 subclones followed during multiple passages of mitotic growth, one instance of a terminal duplication extending into a subtelomeric sequence was observed, but no occurrences of intertelomeric recombination were found. This suggests that intertelomeric recombination is not an important contributor to telomere maintenance in normal K. lactis cells. Rare recombination events resulting in the replacement of a subtelomeric marker with a sequence from another chromosome end also led to the replacement of the telomeric repeat tract. This is consistent with these events being a result of break-induced replication. Movement of a subtelomeric or telomeric sequence from one chromosome end to another was not observed in haploid cells derived from mating and sporulation. This suggests that the exchange of telomeric repeats is not a routine occurrence in K. lactis meiosis.