Institute of Translational Pharmacology, Consiglio Nazionale delle Ricerche (CNR), Roma 00133, Italy.
J Biol Chem. 2019 Aug 16;294(33):12349-12358. doi: 10.1074/jbc.RA119.008529. Epub 2019 Jun 24.
In yeast (), the synthesis of tRNAs by RNA polymerase III (RNAP III) down-regulates the transcription of the nearby RNAP II-transcribed genes by a mechanism that is poorly understood. To clarify the basis of this tRNA gene-mediated (TGM) silencing, here, conducting a bioinformatics analysis of available ChIP-chip and ChIP-sequencing genomic data from yeast, we investigated whether the RNAP III transcriptional machinery can recruit protein factors required for RNAP II transcription. An analysis of 46 genome-wide protein-density profiles revealed that 12 factors normally implicated in RNAP II-mediated gene transcription are more enriched at tRNA than at mRNA loci. These 12 factors typically have RNA-binding properties, participate in the termination stage of the RNAP II transcription, and preferentially localize to the tRNA loci by a mechanism that apparently is based on the RNAP III transcription level. The factors included two kinases of RNAP II (Bur1 and Ctk1), a histone demethylase (Jhd2), and a mutated form of a nucleosome-remodeling factor (Spt6) that have never been reported to be recruited to tRNA loci. Moreover, we show that the expression levels of RNAP II-transcribed genes downstream of tRNA loci correlate with the distance from the tRNA gene by a mechanism that depends on their orientation. These results are consistent with the notion that pre-tRNAs recruit RNAP II-associated factors, thereby reducing the availability of these factors for RNAP II transcription and contributing, at least in part, to the TGM-silencing mechanism.
在酵母中,RNA 聚合酶 III(RNAP III)合成 tRNA 会通过一种机制下调附近由 RNAP II 转录的基因的转录,但这种机制还不太清楚。为了阐明这种 tRNA 基因介导的(TGM)沉默的基础,在这里,我们通过对酵母中可用的 ChIP-chip 和 ChIP-sequencing 基因组数据进行生物信息学分析,研究了 RNAP III 转录机制是否可以招募 RNAP II 转录所需的蛋白质因子。对 46 个全基因组蛋白密度图谱的分析表明,12 个通常与 RNAP II 介导的基因转录有关的因子在 tRNA 上比在 mRNA 上更为丰富。这 12 个因子通常具有 RNA 结合特性,参与 RNAP II 转录的终止阶段,并且通过一种显然基于 RNAP III 转录水平的机制优先定位于 tRNA 位点。这些因子包括两个 RNAP II 的激酶(Bur1 和 Ctk1)、一个组蛋白去甲基酶(Jhd2)和一个核小体重塑因子的突变形式(Spt6),它们从未被报道过被招募到 tRNA 位点。此外,我们还表明,tRNA 位点下游的 RNAP II 转录基因的表达水平与 tRNA 基因的距离相关,其机制依赖于它们的取向。这些结果与前 tRNA 招募 RNAP II 相关因子的观点一致,从而减少了这些因子对 RNAP II 转录的可用性,并在一定程度上促成了 TGM 沉默机制。