Laboratory of tRNA Transcription, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5A, 02-106 Warsaw, Poland.
Wellcome Centre for Cell Biology, The University of Edinburgh, Edinburgh EH8 9YL, UK.
Int J Mol Sci. 2021 Jul 7;22(14):7298. doi: 10.3390/ijms22147298.
The coordinated transcription of the genome is the fundamental mechanism in molecular biology. Transcription in eukaryotes is carried out by three main RNA polymerases: Pol I, II, and III. One basic problem is how a decrease in tRNA levels, by downregulating Pol III efficiency, influences the expression pattern of protein-coding genes. The purpose of this study was to determine the mRNA levels in the yeast mutant and its overdose suppressors, and The mutant prevents assembly of the Pol III complex and functionally mimics similar mutations in human Pol III, which cause hypomyelinating leukodystrophies. We applied RNAseq followed by the hierarchical clustering of our complete RNA-seq transcriptome and functional analysis of genes from the clusters. mRNA upregulation in cells was generally stronger than downregulation. The observed induction of mRNA expression was mostly indirect and resulted from the derepression of general transcription factor Gcn4, differently modulated by suppressor genes. mutation, regardless of the presence of suppressors, also resulted in a weak increase in the expression of ribosome biogenesis genes. mRNA genes that were downregulated by the reduction of Pol III assembly comprise the proteasome complex. In summary, our results provide the regulatory links affected by Pol III assembly that contribute differently to cellular fitness.
基因组的协调转录是分子生物学的基本机制。真核生物中的转录由三种主要的 RNA 聚合酶完成:Pol I、II 和 III。一个基本问题是,tRNA 水平的降低如何通过下调 Pol III 效率来影响蛋白质编码基因的表达模式。本研究的目的是确定酵母突变体及其过表达抑制剂 和 的 mRNA 水平。突变体阻止 Pol III 复合物的组装,并且在功能上模拟了人类 Pol III 中的类似突变,这些突变导致脱髓鞘性白质营养不良。我们应用 RNAseq 并对我们完整的 RNA-seq 转录组进行层次聚类,然后对聚类中的基因进行功能分析。细胞中 mRNA 的上调通常比下调更为强烈。观察到的 mRNA 表达诱导主要是间接的,是由通用转录因子 Gcn4 的去阻遏引起的,而抑制基因对其进行了不同的调节。无论是否存在抑制剂,突变体也导致核糖体生物发生基因的表达微弱增加。Pol III 组装减少下调的 mRNA 基因包括蛋白酶体复合物。总之,我们的结果提供了受 Pol III 组装影响的调节链接,这些链接对细胞适应性有不同的贡献。