The structure and function of transcriptional clusters in bacteria

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