Defining the influence of the A12 subunit on transcription elongation and termination by rna polymerase I. Defining the influence of the A12 subunit on transcription elongation and termination by rna polymerase I

we performed Native Elongating Transcript Sequencing (NET-seq) on an rpa12Δ strain of S. cerevisiae and evaluated the resultant change in Pol I occupancy throughout the 35S gene and the IGS. Compared to WT, we observed template sequence-specific changes in Pol I occupancy throughout the 35S gene. We also observed rpa12Δ Pol I occupancy downstream of both termination sites and throughout most of the IGS, including the 5S gene. Occupancy of rpa12Δ Pol I increased just upstream of the promoter proximal Reb1 binding site and dropped significantly after, implicating this site as a third terminator for Pol I transcription. Overall design: Native Elongating Transcript Sequencing (NET-seq) was performed in triplicate on WT and rpa12Δ S. cerevisiae strains.