Systematic functional and mechanistic analysis of gene regulation by antisense transcription in Saccharomyces cerevisiae

细胞蛋白质的动态平衡确保每种蛋白质在其在生物体内的功能位置处于最佳水平。从转录到蛋白质组周转，细胞机制的多个层次上的一系列调节回路控制着蛋白质的动态平衡。最近的研究揭示了真核生物中普遍存在的非编码转录，其中很大一部分基因与非编码反义转录产物重叠。一些单基因研究表明，这些反义转录本在调节正义基因的表达方面发挥着重要作用。该项目的范围是以酵母为模式生物，通过有针对性地干扰反义转录，从整体上表征反义调控的功能和机制。在面向基因组生物学和生物信息学的小组和面向细胞生物学的小组之间的合作中，我们将应用一种系统的高通量方法来研究反义转录对蛋白质表达水平和细胞特异性表达的影响，使用整个基因组的正义-反义对的全面收集。该项目的目的是通过全球和重点机制研究，利用高含量活细胞显微镜、高通量遗传分析和表型图谱，破译反义转录调控的表型影响、分子机制和功能原理。

Cellular protein homeostasis ensures optimal levels of every protein at the location of its function within the organism. Protein homeostasis is governed by a series of regulatory circuits at multiple levels of the cellular machinery, from transcription to proteome turnover. Recent studies have revealed the existence of pervasive non-coding transcription in eukaryotes, whereby a large fraction of genes overlap with non-coding antisense transcripts. Several single-gene studies have demonstrated that these antisense transcripts play significant roles in regulating expression of the sense gene. The scope of this project is to globally characterize the functions and mechanisms of antisense-mediated regulation, via targeted disruptions of antisense transcription, using yeast as a model organism. In a collaboration between a genome biology and bioinformatics oriented group and a cell biology oriented group, we will apply a systematic high-throughput approach to study the impact of antisense transcription on protein expression levels and cell-specific expression, using a comprehensive collection of sense-antisense pairs across the genome. The aim of the project is to decipher the phenotypic impact, molecular mechanisms and functional rationale of antisense transcriptional control by means of both global and focused mechanistic studies, using high content live cell microscopy, high-throughput genetic analysis, and phenotypic profiling.