Genome-wide profiling of Scc1 subunit in wild type cells and in cells in which chromatin structure is perturbed (histone H4 depletion).

The main objective of this study is to determine how chromatin assembly during DNA replication influences cohesin deposition Cohesin deposition was determined by ChIP-on-ChIP in 2 different genetic backgrounds (wild-type and t::HHF2-W303 bar1∆ background) that carried a tagged version of SCC1 (SCC1-6HA). t::HHF2 is a genetic background in which both loci that codify histone H4 are deleted and all histone H4 is expressed from a doxycycline regulatable promoter (tetON). Histone depletion was induced in G1 arrested cells changing the concentration of doxycicline (5 µg/ml ≈ wild type to 0.25 µg/ml = histone depletion). 2 samples for each genetic background (2 independent experiments) were collected at the G2/M transition. Each experiment carried a non-tagged version of the wild type strain. One of them was analyzed to determine all non-specific signal.