Role and regulation of convergent antisense transcription by RNA polymerase II

RNA 聚合酶 II 聚合反义转录的作用和调节

• 批准号: 418415292
• 负责人: Dr. Andreas Mayer
• 金额: --
• 依托单位: Max-Planck-Institut für molekulare Genetik (MPIMG)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/418415292?language=en
• 关键词: Role; regulation; convergent; antisense; transcription

Gene transcription by RNA polymerase II (Pol II) is not a unidirectional process. Transcription in the sense direction is usually accompanied by antisense transcription in the opposite orientation. Antisense transcription is emerging as a new layer in regulating the expression of the sense gene. We have recently developed native elongating transcript sequencing (NET-seq) as a quantitative genome-wide approach for studying the full spectrum of sense and antisense transcription with nucleotide precision in human cells (Mayer et al., Cell, 2015; Mayer et al., Nat Protocols, 2016). Application of this high-resolution method has revealed a new antisense transcriptional activity of Pol II in the promoter-proximal region of a large set of protein-coding and long non-coding RNA genes, called convergent antisense transcription. Convergent antisense transcription predominantly occurs at lower expressed genes and inversely correlates with sense transcription suggesting a potential regulatory link. However, key aspects of convergent antisense transcription remain undefined. Here, we propose to address two fundamental questions: (1) What molecular mechanisms drive this antisense transcriptional activity? (2) What is the functional role of convergent antisense transcription in controlling the transcriptional output of cells? To understand the determinants and functions of convergent antisense transcription in human cells, we will employ an interdisciplinary combination of high-resolution genome-wide approaches, genome editing tools and single-molecule microscopy.

RNA 聚合酶 II (Pol II) 的基因转录不是单向过程。有义方向的转录通常伴随着相反方向的反义转录。反义转录正在成为调节有义基因表达的新层。我们最近开发了天然延伸转录本测序 (NET-seq)，作为一种全基因组定量方法，用于研究人类细胞中核苷酸精度的全谱有义和反义转录（Mayer 等人，Cell，2015；Mayer 等人，Nat Protocols，2016）。这种高分辨率方法的应用揭示了 Pol II 在一大组蛋白质编码和长非编码 RNA 基因的启动子近端区域中的新反义转录活性，称为聚合反义转录。趋同反义转录主要发生在表达较低的基因上，并与有义转录呈负相关，表明存在潜在的调控联系。然而，收敛反义转录的关键方面仍不清楚。在这里，我们建议解决两个基本问题：（1）什么分子机制驱动这种反义转录活性？ (2) 收敛反义转录在控制细胞转录输出中的功能作用是什么？为了了解人类细胞中聚合反义转录的决定因素和功能，我们将采用高分辨率全基因组方法、基因组编辑工具和单分子显微镜的跨学科组合。