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. 2021 Dec 30;50(2):899–914. doi: 10.1093/nar/gkab1253

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — The ade6-3101 hotspot manifests Tel1-dependent DSB interference; Tel1-independent DSB competition indicates separate mechanisms. rad50S strains were induced for meiosis, and DNA analyzed as in Figure 3, except meiosis was at 25°C in (B). (A) Both ade6-3049 and ade6-3101 manifest Tel1-dependent DSB interference. DNA was digested with NotI and analyzed on four to six blots with a probe between ade6 and the 75R DSB hotspot (left panel) or between ade6 and the tel1L hotspot near tel1 (right panel). Double-cut DSBs (black arrows; 75 kb left and 40 kb right) were evident in tel1Δ (left and middle lane sets) but not in tel1⁺ (right lane set). Coefficients of coincidence (CoC; mean ± SEM) show positive DSB interference (1 - CoC) in tel1⁺ and negative interference in tel1Δ. Single-cut DSBs and frequencies are visible in Supplementary Figure S3 using a different radioactive probe. (B) DSB competition at mbs1 is Tel1-independent. DNA was digested with NotI and analyzed on three to seven blots with a probe at the left end of the 501 kb NotI fragment J. DSBs at both mbs1⁺ and ura1::hph hotspots were reduced in the presence of the other hotspot (compare the double hotspot in the middle lane set to either single hotspot; **P = 0.007 for mbs1⁺ and **P = 0.0034 for ura1::hph), indicating mutual DSB competition. This competition was also present without Tel1 (right panel; **P = 0.002 for mbs1⁺ and *P = 0.018 for ura1::hph).