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. Triplicate NET-seq libraries of wild-type (WT) yeast and rpa190-F1205H yeast were prepared and analyzed.

本研究旨在利用携带RNA聚合酶I（Pol I）突变的酵母菌株，在体内探究Pol I转录延伸速率与核糖体RNA加工之间的关联。该突变体rpa190-F1205H此前已被表征为体外转录延伸速率降低。本研究采用原生延伸转录物测序（native elongating transcript sequencing, NET-seq）技术，测定该突变对体内Pol I占据分布的影响。本研究结果表明，该突变可加剧Pol I的暂停现象，尤其集中于35S基因的前半段区域，并导致Pol I的占据分布出现序列特异性改变。我们制备并分析了野生型（WT）酵母与rpa190-F1205H突变酵母的三份重复NET-seq文库。