Exploring the dynamics of DNA repair in an intracellular pathogen using single molecule tracking

Abstract
Macrophages are a key cell type of the innate immune system which kill bacteria following uptake by phagocytosis. Some bacterial species known as 'intracellular pathogens' are able to avoid killing by the phagocytic pathway. For example, Salmonella enterica serovar Typhimurium (S. Tm) is able to survive and replicate within its host cell by modifying macrophage function. Following phagocytosis, the macrophage produces reactive oxygen species (ROS), which cause damage to bacterial DNA. However, the nature of DNA damage repair this elicits in intracellular pathogens such as S. Tm, and the consequences of this for S. Tm residing in the macrophage, is not understood. Single molecule tracking (SMT) is a technique which enables the localisation and tracking of spatially isolated molecules at super-resolution, in real-time. This project aims to use SMT to study the activity of DNA damage repair proteins in S. Tm within macrophages. This will facilitate an understanding of which DNA repair pathways become active in S. Tm in relation to ROS produced by the macrophage, and how this ultimately affects the fate of heterogeneous S. Tm populations. Together, this will provide us with novel insights regarding the complex host-pathogen interactions occurring during intracellular S. Tm infection.
BBSRC Priority Areas
Transformative technologies, understanding the rules of life (advancing the frontiers of bioscience discovery).