Dependent around the SC element Zip1 [16, 17] and a few specifications regarding the regulation

Dependent around the SC element Zip1 [16, 17] and a few specifications regarding the regulation of complete centromere coupling have started to emerge, like roles for the meiotic cohesin Rec8 [22], for the SC element Zip3 in coupling and tethering [16, 23], and for the phosphorylation of Zip1 by ATM/ATR DSB checkpoint kinases [18]. On the other hand, the underlying architecture of centromere coupling remains to become understood. In specific, the presence of an interaction pattern of centromeres, if any, may point towards an intrinsic mechanism for coupling. So far prior research have relied on low-scale, standard approaches not amenable to testing this hypothesis on a larger level. The budding yeast genome, regardless of its compact size, exhibits a high level of inter-chromosomal contacts and long-range cis interactions among distant loci [24]. Chromosome Conformation Capture (3C) enables the detection of DNA regions in close nuclear proximity by means of formaldehyde crosslinking of such interactions followed by restriction enzyme digestion, dilute ligation to favor intra-molecular items that are crosslinked, and PCR detection [25]. 3C was first developed in budding yeast to study chromosome dynamics through meiosis and higherorder chromatin organization [25], and has since been applied the investigation of diverse biological processes for instance silencing [26], organization on the pericentric chromatin [27], and gene looping [28, 29]. 3C has yielded various associated procedures which have enabled the characterization of long-range genome associations in mammals [304]. One particular such variant, Taqmanbased 3C-qPCR, is well suited for focused studies, with higher sensitivity and dynamic range, low background and quantitative detection of Bepotastine Antagonist interacting fragments [32]. Right here we present the very first numerous pairwise characterization of centromere coupling. We modified and combined the yeast 3C protocol [35, 36] with Taqman-based real-time detection of 3C ligation products (3C-qPCR) [32] to quantify all probable non-homologous interactions in between the 16 centromeres (CENs) of S. cerevisiae during meiosis. We observed a non-random CEN interaction pattern determined by similarity of chromosome sizes in strains capable of coupling (spo11 diploids and haploids), which is Share this post on: