4tU RNA-seq from wild-type and Rpb7-QHF cells
Our structural and biochemical studies show that S. cerevisiae RNA Polymerase II (RNAPII) homodimerizes through the stalk domain (formed by the Rpb4-Rpb7 subunits). To explore the biological impact of disrupting this interaction we introduced a triple point mutation at the RNAPII dimerization interface in Rpb7 (Q96A, H97A, F109K). To test the impact of the dimerization mutant on RNA synthesis and 3' end processing in WT and Rpb7-QHF cells, we labelled nascent transcripts with 4tU which enabled subsequent biotinylation and purification of newly-synthesized transcripts. Nascent transcripts were then used to prepare strand-specific RNA-seq libraries.