Bringing Rad52 foci into focus

The Rad52 protein has pivotal functions in double strand break repair and homologous recombination. The activity of Rad52 is often monitored by the subnuclear foci that it forms spontaneously in S phase or after DNA damage (Lisby et al., 2001). In mammals, the functions of yeast Rad52 may be divided between human RAD52 and the tumor suppressor BRCA2 (Feng et al., 2011). The full host of molecular players that govern Rad52 focus formation and maintenance was not well known when we initiated our screen. Using a high-content, image-based assay, we assessed the proportion of cells containing spontaneous Rad52-YFP foci in 4,805 viable Saccharomyces cerevisiae deletion strains (Alvaro et al., 2007). Starting with 96-well arrays of a deletion strain library , we created hybrid diploid strains (homozygous for the deletions) using systematic hybrid loss of heterozygosity (SHyLOH; Alvaro et al., 2006). We then manually and sequentially examined each strain using epifluorescence micros-copy for the presence of Rad52-YFP foci. All of our image analysis was performed manually. As is often the case, our screen was published showing only a couple of representative images and providing data tables to summarize the findings. Tomes of data that could not be included in the published paper were relegated to supplemental Excel tables, typical of genome-wide screens. Also, the raw image data were sequestered in the laboratory on DVDs. With considerable help from JCB and Glencoe In 2007, we published the results of a genome-wide screen for ORFs that affect the frequency of Rad52 foci in yeast. That paper was published within the constraints of conventional online publishing tools, and it provided only a glimpse into the actual screen data. New tools in the JCB DataViewer now show how these data can—and should—be shared. Software, we are delighted that the raw data from our Rad52 screen are now freely available online through the JCB Data-Viewer. A new interface within the JCB DataViewer brings presentation and preservation of high-content, multidimen-sional image-based screening data to a whole new level. To facilitate the development of this new interface, JCB required a dataset that was not time sensitive, and we were happy to provide our previously published Rad52 data. In the future , this new interface will be used to present high-content screening (HCS) datasets linked to published JCB papers. Indeed, the first publication of this sort appears in this issue of JCB (Rohn et al., 2011). The presentation of …