Department of Biology, Indiana University, Bloomington, Indiana, USA.
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, France.
Mol Cell Biol. 2018 Nov 28;38(24). doi: 10.1128/MCB.00296-18. Print 2018 Dec 15.
Eukaryotic RNA polymerase II (RNAPII) transcribes mRNA genes and non-protein-coding RNA (ncRNA) genes, including those encoding small nuclear and nucleolar RNAs (sn/snoRNAs). In metazoans, RNAPII transcription of sn/snoRNAs is facilitated by a number of specialized complexes, but no such complexes have been discovered in yeast. It has been proposed that yeast sn/snoRNA and mRNA expression relies on a set of common factors, but the extent to which regulators of mRNA genes function at yeast sn/snoRNA genes is unclear. Here, we investigated a potential role for the Mediator complex, essential for mRNA gene transcription, in sn/snoRNA gene transcription. We found that Mediator maps to sn/snoRNA gene regulatory regions and that rapid depletion of the essential structural subunit Med14 strongly reduces RNAPII and TFIIB occupancy as well as nascent transcription of sn/snoRNA genes. Deletion of Med3 and Med15, subunits of the activator-interacting Mediator tail module, does not affect Mediator recruitment to or RNAPII and TFIIB occupancy of sn/snoRNA genes. Our analyses suggest that Mediator promotes PIC formation and transcription at sn/snoRNA genes, expanding the role of this critical regulator beyond its known functions in mRNA gene transcription and demonstrating further mechanistic similarity between the transcription of mRNA and sn/snoRNA genes.
真核生物 RNA 聚合酶 II(RNAPII)转录 mRNA 基因和非蛋白编码 RNA(ncRNA)基因,包括编码小核和核仁 RNA(sn/snoRNA)的基因。在真核生物中,许多专门的复合物促进了 sn/snoRNA 的 RNAPII 转录,但在酵母中尚未发现此类复合物。有人提出,酵母 sn/snoRNA 和 mRNA 的表达依赖于一组共同的因子,但调控 mRNA 基因的因子在多大程度上作用于酵母 sn/snoRNA 基因尚不清楚。在这里,我们研究了中介复合物(essential for mRNA gene transcription)在 sn/snoRNA 基因转录中的潜在作用。我们发现中介复合物定位于 sn/snoRNA 基因调控区,并且必需的结构亚基 Med14 的快速耗尽强烈降低了 RNAPII 和 TFIIB 的占有率以及 sn/snoRNA 基因的新生转录。删除激活子相互作用的中介尾部模块的亚基 Med3 和 Med15 并不影响中介复合物对 sn/snoRNA 基因的招募或 RNAPII 和 TFIIB 的占有率。我们的分析表明,中介复合物促进了 PIC 在 sn/snoRNA 基因上的形成和转录,将这一关键调控因子的作用扩展到其在 mRNA 基因转录中的已知功能之外,并进一步证明了 mRNA 和 sn/snoRNA 基因转录之间的机制相似性。