Mesoscale structure in metabolism and cell death

Abstract

The emerging field of bacterial cell biology is beginning to shed light on the grand subcellular complexity of these small cells. Advances in imaging technologies and fluorescent proteins have peeled back the veil that had left the subcellular organization of bacteria largely unappreciated. Here I take these technologies further to demonstrate the importance of mesoscale architecture in bacteria in two fundamental processes: the regulation of metabolism and the phenotypic classification of cell death.

For its us in metabolism I forward-engineered mesoscale structures to demonstrate their feasibility in controlling metabolic flux based on a model of the theoretically optimal enzyme cluster. I then designed and validated a set of plug-and-play clustering tags for widespread actualization of enzyme clustering in metabolic engineering. Using a technique known as bacterial cytological profiling (BCP) I utilize various mesoscale structures as classification features to find the mechanism of action (MOA) of an ecologically important antibiotic, tropodithietic acid (TDA). Finding TDA's MOA suggests a mechanism of resistance in its bacterial producer as well as a potential anticancer property for TDA. I then scaled BCP for high-throughput applications and created a more informative analysis pipeline that I used to assess a panel of molecules with previously unknown mechanisms of action. This generated 8 antibiotic lead compounds with potentially novel MOAs.