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 rpa12? 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 rpa12? Pol I occupancy downstream of both termination sites and throughout most of the IGS, including the 5S gene. Occupancy of rpa12? 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 rpa12? S. cerevisiae strains.

我们对酿酒酵母（S. cerevisiae）的rpa12?菌株开展了原生延伸转录测序（Native Elongating Transcript Sequencing, NET-seq），并评估了由此导致的RNA聚合酶I（Pol I）在35S基因与基因间区（IGS）全区域的结合占有率变化。与野生型（Wild Type, WT）菌株相比，我们观测到RNA聚合酶I在35S基因全区域的结合占有率存在模板序列特异性改变。此外，我们还发现rpa12?菌株的RNA聚合酶I在两个终止位点下游以及基因间区的大部分区域（包括5S基因）均出现结合占有率异常。具体来看，rpa12?菌株的RNA聚合酶I在启动子近端的Reb1结合位点上游的结合占有率升高，而在该位点之后结合占有率显著下降，这提示该位点可作为RNA聚合酶I转录的第三个终止子。实验设计：我们对野生型与rpa12?酿酒酵母菌株分别进行了三次生物学重复的原生延伸转录测序。