CRAC of yeast RNA polymerase II and RNases H in asynchronous, G1-arrested, and S-phase cells.

We obtained RNA polymerase II and RNases H occupancy profiles across the genome of S.cerevisiae strains: Sen1 depleted cells using the degron system (including no addition of IAA); sen1-3, RNases H deleted, and wild-type cells. This allowed us to compare RNAPII and RNases H occupancy profiles in different genetic context and/or phases of the cell cycle. The nascent RNA was crosslinked to Rpb1 (RNAPII) or to RNases H (Rnh1 or Rnh201) and sequenced after purification of the target protein under stringent and denaturing conditions. This reveals the position of the target protein at the nucleotide resolution level.