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. 2014 Oct 16;10(10):e1004708. doi: 10.1371/journal.pgen.1004708

Figure 6. Effects of Tpz1-Poz1 interaction disruption mutations on telomere maintenance.

Figure 6

(A) Southern blot analysis and (B) pulsed-field gel analysis for indicated Tpz1-Poz1 interaction disruption mutants. Haploid cells were generated by dissection of spores derived from heterozygous tpz1+/mutated tpz1 diploid cells, and restreaked 5 times on plates prior to preparation of genomic DNA to achieve steady state telomere length, except for tpz1-W499R,I501R poz1Δ and ccq1Δ poz1Δ cells where DNA from survivors with circular chromosomes were made after restreaked twice on plates. For each round of restreak, several faster growing colonies were combined and streaked for single colonies on YES plates. (C) Southern blot analysis and (D) pulsed-field gel analysis for mutants that disrupted either Tpz1-Ccq1 or Tpz1-Poz1 interaction, or both Tpz1-Ccq1 and Tpz1-Poz1 interactions. DNA samples were prepared after restreaked twice on plates. Simultaneous loss of Tpz1-Ccq1 and Tpz1-Poz1 interactions resulted in complete loss of telomeres and circularization of chromosomes, much like in ccq1Δ poz1Δ, tpz1-L449R poz1Δ and tpz1-W498R,I501R ccq1Δ cells.