Mutations in yeast Pcf11, a conserved protein essential for mRNA 3′ end processing and transcription termination, elicit the Environmental Stress Response
Mutations in yeast Pcf11, a conserved protein essential for mRNA 3′ end processing and transcription termination, elicit the Environmental Stress Response
复制标题
DOI：
10.1093/genetics/iyad199
复制
发表时间：
2023-11-15
期刊：
影响因子：
3.3
通讯作者：
Moore，Claire
中科院分区：
文献类型：
--
作者：
Graber，Joel H.;Hoskinson，Derick;Moore，Claire
Pcf 11蛋白是切割和聚腺苷酸化真核mRNA前体的大复合物的必需亚基。它还与基因循环、RNA聚合酶II（Pol II）转录物的终止和mRNA输出功能相关。我们研究了酵母菌Pcf 11的一个特征不明确但保守的结构域（氨基酸142-225），发现虽然它不是mRNA 3′端加工或蛋白编码基因poly（A）位点下游终止所必需的，但它的存在改善了与Pol II的相互作用以及基因启动子附近转录终止子的使用。全基因组Pol II占用与Pcf 11缺少这个区域的细胞，以及Pcf 11突变的Pol II CTD相互作用结构域的分析表明，mRNA表达的系统性变化主要是在转录水平介导的。全球表达分析还表明，一般的应激反应，包括激活和抑制的特定基因集已知的调节，在响应各种各样的压力，诱导在两个opcf 11突变体，即使细胞生长在最佳条件。这些突变体还导致细胞壁相关基因的不平衡表达，这些基因不激活细胞壁完整性途径，但与强烈的咖啡因敏感性有关。基于这些发现，我们提出Pcf 11可以调节特定功能组基因的表达水平，而不涉及其在mRNA 3′端加工中的作用。
The Pcf11 protein is an essential subunit of the large complex that cleaves and polyadenylates eukaryotic mRNA precursor. It has also been functionally linked to gene-looping, termination of RNA Polymerase II (Pol II) transcripts, and mRNA export. We have examined a poorly characterized but conserved domain (amino acids 142–225) of theSaccharomyces cerevisiaePcf11 and found that while it is not needed for mRNA 3′ end processing or termination downstream of the poly(A) sites of protein-coding genes, its presence improves the interaction with Pol II and the use of transcription terminators near gene promoters. Analysis of genome-wide Pol II occupancy in cells with Pcf11 missing this region, as well as Pcf11 mutated in the Pol II CTD Interacting Domain, indicates that systematic changes in mRNA expression are mediated primarily at the level of transcription. Global expression analysis also shows that a general stress response, involving both activation and suppression of specific gene sets known to be regulated in response to a wide variety of stresses, is induced in the twopcf11mutants, even though cells are grown in optimal conditions. The mutants also cause an unbalanced expression of cell wall-related genes that does not activate the Cell Wall Integrity pathway but is associated with strong caffeine sensitivity. Based on these findings, we propose that Pcf11 can modulate the expression level of specific functional groups of genes in ways that do not involve its well-characterized role in mRNA 3′ end processing.