Analysis by Mnase-seq of the chromatin structure of wild type cells and cells in which chromatin structure is perturbed (histone H4 depletion) in the presence or absence of the kleisin subunit of cohesin, Scc1

The main objective of this study is to determine how cohesin influences chromatin structure and chromatin assembly during DNA replication Overall design: Chromatin structure has been determined by MNase-seq in 4 different genetic backgrounds (wild-type, scc1-73, tHHF2 and tHHF2 scc1-73) in S.cerevisiae cells (W303 bar1? background). scc1-73 is a thermosensitive allele of the kleisin subunit of the mitotic cohesin complex that was used to analyze the effects of cohesin inactivation on chromatin. tHHF2 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.