Zhang Yinfeng, French Sarah L, Beyer Ann L, Schneider David A
From the Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, Alabama 35294-0024 and.
the Department of Microbiology, University of Virginia, Charlottesville, Virginia 22908.
J Biol Chem. 2016 Feb 5;291(6):3010-8. doi: 10.1074/jbc.M115.673442. Epub 2015 Dec 9.
Although ribosomal RNA represents the majority of cellular RNA, and ribosome synthesis is closely connected to cell growth and proliferation rates, a complete understanding of the factors that influence transcription of ribosomal DNA is lacking. Here, we show that the THO complex positively affects transcription by RNA polymerase I (Pol I). We found that THO physically associates with the rDNA repeat and interacts genetically with Pol I transcription initiation factors. Pol I transcription in hpr1 or tho2 null mutants is dramatically reduced to less than 20% of the WT level. Pol I occupancy of the coding region of the rDNA in THO mutants is decreased to ~50% of WT level. Furthermore, although the percentage of active rDNA repeats remains unaffected in the mutant cells, the overall rDNA copy number increases ~2-fold compared with WT. Together, these data show that perturbation of THO function impairs transcription initiation and elongation by Pol I, identifying a new cellular target for the conserved THO complex.
尽管核糖体RNA占细胞RNA的大部分,且核糖体合成与细胞生长和增殖速率密切相关,但目前仍缺乏对影响核糖体DNA转录因素的全面了解。在此,我们表明THO复合物对RNA聚合酶I(Pol I)的转录具有正向影响。我们发现THO与rDNA重复序列存在物理关联,并与Pol I转录起始因子存在遗传相互作用。hpr1或tho2基因敲除突变体中的Pol I转录显著降低至野生型水平的不到20%。THO突变体中rDNA编码区的Pol I占据率降至野生型水平的约50%。此外,尽管突变细胞中活跃rDNA重复序列的百分比未受影响,但与野生型相比,总体rDNA拷贝数增加了约2倍。这些数据共同表明,THO功能的扰动会损害Pol I的转录起始和延伸,从而确定了保守的THO复合物的一个新的细胞靶点。