Collisions of replication and transcription machinery in Escherichia coli.

DNA duplication is essential for life. Numerous proteins bind and interact with DNA. When replication forks collide with proteins bound to DNA they can be blocked and this causes problems for genome stability. In bacteria DNA replication occurs at the same time as transcription of genes so the most common obstacles for replication forks are transcribing RNA polymerases. Understanding how the simultaneous processes of replication and transcription interfere with each other is a key challenge for molecular microbiology and more broadly molecular biology.

This project will investigate replisome-transcription machinery conflicts in Escherichia coli using a combination of biochemical, biophysical and microscopy-based approaches. The aim is to uncover the mechanistic detail of replication and transcription collisions and shed light on the systems involved in preventing them. Bacterial replication can be reconstituted in a tube and we can use this to biochemically investigate replication blocks and their consequences. This project will also characterise the location and interaction-partners of proteins of interest using advanced microscopy techniques and biophysical methods. The combination of biophysics, cutting edge imaging, biochemistry, genetics and molecular biology is a unique synergy of approaches that will ensure a highly successful project with training in pioneering techniques.