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. 2018 Apr 1;32(7-8):537–554. doi: 10.1101/gad.311712.118

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© 2018 Begnis et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 5. — RNAi-dependent pathways confer illegitimate rDNA translocation. (A) Schematic of HAATI^rDNA+Trt1 cells forced into subsequent Trt1 loss. (B) Table summarizing the frequency of survivor formation under noncompetitive (single-colony) growth conditions. Single-colony data for “rDNA on chromosome III” (i.e., wild-type cells), taken from Figure 4D, are shown for comparison. (C) PFGE analysis of whole chromosomes hybridized with rDNA probe. Cultures of the indicated genotypes were propagated in liquid for 5 d. Aliquots were sampled at days 1, 3, and 5 for transformation with a plasmid encoding trt1⁺, which confers telomere addition and gel entry and thus a snapshot of chromosome length over time in the HAATI^rDNA population. (D) Levels of H3K9me enrichment in wild-type and dcr1Δ (trt1⁺) cells before (+EtOH), during (+TSA [trichostatin A]), and after (+REC, for recovery) TSA treatment. Levels of the indicated genome regions in H3K9me2 immunoprecipitates were quantified by RT–PCR using primers for pericentromeric dg/dh repeats, subtelomeric (subTELO) regions, and the 18S rDNA. Means are shown; error bars represent standard deviations of three biological repeats. (E) Rad51 is essential for HAATI maintenance. Fivefold serial dilutions of the indicated strains on medium selecting for (FOA) or against (−URA) loss of plasmid expressing Rad51 (“+p51”). (F) Rad52 is essential for HAATI maintenance. Fivefold serial dilutions of the indicated strains on medium selecting for (FOA) or against (−URA) loss of plasmid expressing Rad52 (“+p52”).