Impairment of RNA polymerase I elongation rate induces defects in ribosomal RNA processing and ribosome biogenesis

The objective of this study was to investigate the relationship between Pol I transcription elongation rate and ribosomal RNA processing in vivo by using a yeast strain containing a mutation in Pol I. This mutation, rpa190-F1205H, has been previously characterized to have a reduced elongation rate in vitro. Here, we used native elongating transcript sequencing (NET-seq) to determine the effect of this mutation on Pol I occupancy in vivo. Our findings demonstrate that this mutation induces increased Pol I pausing, especially in the first half of the 35S gene, and causes sequence-specific changes in Pol I occupancy. Overall design: Triplicate NET-seq libraries of wild-type (WT) yeast and rpa190-F1205H yeast were prepared and analyzed.