The structure and function of transcriptional clusters in bacteria

在转录过程中，DNA序列被复制成RNA分子。它们被用来产生重要的细胞成分，如蛋白质和核糖体，使转录成为所有生命的基础。细菌利用它来对环境做出反应，在不同的条件下转录不同的分子。最近超分辨率显微镜的发展使我们能够在一个新的细节水平上研究这一点，揭示了在快速生长的细菌中形成的RNA聚合酶（RNAP）大簇，即负责转录的蛋白质。然而，关于集群如何形成、它们的功能和它们的组成仍然存在不确定性。该项目将通过收集活细胞中的超分辨率显微镜数据和处理所需的复杂数据分析来回答这些问题。我们将确定簇是否通过相分离形成，就像水中的油滴；首先，通过观察活细胞中的集群，然后压缩单个细胞。接下来，我们将评估簇的功能是否通过跟踪单个分子的扩散来加速RNAP对DNA靶标的搜索。最后，我们将通过分析蛋白质聚类和扩散来研究哪些蛋白质形成簇的一部分。这些见解将提高我们对转录的基本理解，并可用于对抗抗生素耐药性和推进生物技术进程。BBSRC解决的优先领域-理解生活规则-变革技术

In transcription, DNA sequences are copied to make RNA molecules. These are used to produce vital cellular components such as proteins and ribosomes, making transcription fundamental for all life. Bacteria use this to respond to their environment, transcribing different molecules in response to different conditions. The recent development of super-resolution microscopy has enabled us to investigate this in a new level of detail, revealing that large clusters of RNA polymerase (RNAP), the protein responsible for transcription, form in fast-growing bacteria. However, uncertainties remain over how clusters form, their function, and their components. This project will answer these questions by gathering super-resolution microscopy data in live cells and tackling the complex data analysis required. We will determine whether clusters form via phase separation, like oil droplets in water; firstly, by observing clusters in live cells, then compressing individual cells. Next, we will assess whether the function of clusters is to accelerate RNAP's search for DNA targets by tracking the diffusion of single molecules. Finally, we will investigate which proteins form part of clusters by analysing protein clustering and diffusion. These insights will improve our fundamental understanding of transcription and could be leveraged to fight antibiotic resistance and advance biotechnology processes.BBSRC priority areas addressed - Understanding the rules of life - Transformative technologies