FIG 3.

The SiRTA Stim and Core sequences are sufficient to stimulate de novo telomere addition at an ectopic location. (A) Top schematic, sequence of SiRTA 5L-35 as described for Fig. 2A. Bottom schematic, spacerΔ mutation created in SiRTA 5L-35. The dashed line indicates deleted nucleotides. (B) The absolute frequency (% total cells) of GCR formation at SiRTA 5L-35 is shown for the spacerΔ variant at its endogenous location on chromosome V and at an ectopic site on chromosome VII. Data for WT SiRTA 5L-35 are shown for comparison (same as Fig. 2B). Values are the averages from three independent experiments with standard deviations. (C) Schematic of the modified left arm of chromosome VII. Sizes of the regions between the integrated spacerΔ sequence and either the HO cleavage site (telomere proximal) or the most distal essential gene (BRR6; centromere proximal) are indicated. (D) The percentage of GCR events occurring in each indicated region on chromosome VII is shown for the experimental strain (SiRTA 5L-35 spacerΔ) and a control strain (no integration). In the control strain, no GCR events were observed in the 219-bp region that is replaced by the spacerΔ variant in the experimental strain. Values are averages from three independent experiments with standard deviations.