Fig. 2: Rif2 BAT motif inhibits NHEJ at broken ends.

A I-SceI assay used to estimate NHEJ efficiency. Two inverted I-SceI sites are inserted at the endogenous URA3 gene. Most survivors to continuous I-SceI expression have eliminated the I-SceI sites by fusing the distal broken ends²¹. B NHEJ inhibition by Rap1 C-terminal domain and Rif2 N-terminal region targeted at broken ends (lif1∆: NHEJ-deficient cells). Means from independent cell cultures. C Increasing distances between the broken end and the Gal4 binding sites decrease NHEJ inhibition by Rif2. D Rif2 N-terminal truncations impacting the ability to inhibit NHEJ at broken ends. Gal4_DBD and Gal4_DBD-Rif2_fragments expressed from a centromeric plasmid. E Rif2 mutations impacting its ability to inhibit NHEJ at broken ends. Gal4_DBD and Gal4_DBD-Rif2_1-60 expressed from an integrated plasmid. Gal4_DBD-Rif2_1-395 expressed from a centromeric plasmid. F The rif2-F8A mutation exposes telomeres to NHEJ in cells lacking Sir4 (fusions between X and Y′ telomeres). Experiment reproduced three times. G K. lactis Orc4 expression complements Rif2 loss for NHEJ inhibition by Rap1 C-terminal domain in S. cerevisiae.