Rad52-C180A-CFP Is Recruited to a Specific DNA Double-Strand Break

<div><p>(A) Assay in Chromosome III for visualizing an HO-endonuclease inducible DSB. Yellow boxes: <i>lacO</i> sites. Cyan triangle: HO cut-site (HOcs). Solid circle: centromere.</p><p>(B) Localization of a Rad52–CFP focus to an HO-endonuclease-induced DSB. The panels show YFP, CFP, CFP/YFP-merged, and DIC images of representative cells of a strain expressing Rad52-C180A-CFP in a strain with a <i>lacO</i> tandem array next to an HOcs on Chromosome III (left, strain W4021-20A) and in a strain with an HOcs on Chromosome III and a <i>lacO</i> tandem array on Chromosome IV (right, strain W4341-16A). The <i>lacO</i> tandem array is visualized by LacI-YFP as a yellow focus. The marked foci (arrowheads) in the left panels are examples of Rad52/LacI co-localization and the arrowheads in the example on the right indicate the absence of co-localization when the <i>lacO</i> elements and the HOcs are on different chromosomes. Scale bar, 3 μm.</p><p>(C) Quantitative analysis of co-localization between Rad52-CFP and YFP-LacI foci. Chromosomal locations of the HOcs and the <i>lacO</i> tandem array are given below the histogram columns. As a control, strain W4341-6D with no HOcs was analyzed. The wild-type dataset shown for comparison is from [<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.0020194#pgen-0020194-b039" target="_blank">39</a>].</p>