In budding yeast, the 3' end processing of mRNA and the coupled termination of transcription by RNAPII requires the CF IA complex. We have earlier demonstrated a role for the Clp1 subunit of this complex in termination and promoter-associated transcription of CHA1. To assess the generality of the observed function of Clp1 in transcription, we tested the effect of Clp1 on transcription on a genomewide scale using the Global Run-On-Seq (GRO-Seq) approach. GRO-Seq analysis showed the polymerase reading through the termination signal in the downstream region of highly transcribed genes in a temperature-sensitive mutant of Clp1 at elevated temperature. No such terminator readthrough was observed in the mutant at the permissive temperature. The poly(A)-independent termination of transcription of snoRNAs, however, remained unaffected in the absence of Clp1 activity. These results strongly suggest a role for Clp1 in poly(A)-coupled termination of transcription. Furthermore, the density of antisense transcribing polymerase upstream of the promoter region exhibited an increase in the absence of Clp1 activity, thus implicating Clp1 in promoter directionality. The overall conclusion of these results is that Clp1 plays a general role in poly(A)-coupled termination of RNAPII transcription and in enhancing promoter directionality in budding yeast.

中文翻译：

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Clp1功能在芽酵母中转录的全基因组分析。

在发芽酵母中，mRNA的3'末端加工和RNAPII转录的偶联终止需要CF IA复合物。我们之前已经证明了该复合物的Clp1亚基在CHA1终止和启动子相关转录中的作用。为了评估Clp1在转录中功能的一般性，我们使用Global Run-On-Seq（GRO-Seq）方法在全基因组范围内测试了Clp1对转录的影响。GRO-Seq分析显示，聚合酶通过高温下Clp1的温度敏感突变体中高转录基因下游区域的终止信号读取。在允许的温度下，在突变体中未观察到这样的终止子通读。但是，snoRNA的poly（A）依赖性终止转录 在没有Clp1活性的情况下仍不受影响。这些结果强烈暗示了Clp1在poly（A）偶联的转录终止中的作用。此外，启动子区域上游的反义转录聚合酶的密度在没有Clp1活性的情况下表现出增加，因此暗示Clp1具有启动子方向性。这些结果的总体结论是，Clp1在poly（A）偶联的RNAPII转录终止和增强发芽酵母中的启动子方向性中起一般作用。