Mechanism and implications of ABC-F-mediated antibiotic resistance in clinically relevant bacteria

We will extend our live-cell single-molecule tracking techniques to study how ABC-F proteins mediate antibiotic resistance in clinically relevant bacteria.

In particular, we will study the dynamic interactions of ABC-Fs with the bacterial protein synthesis machinery, and evaluate predictions from the recently proposed mechanistic model.A postdoctoral fellow and an experienced researcher will fluorescence label ABC-Fs (Years 1-2) and study their interaction with the protein synthesis machinery directly inside relevant living bacterial cells (Years 1-3).

This will require new technical approaches, including introduction of test-tube fluorescence-labeled proteins into bacteria by electroporation.

They will also use our existing approaches, e.g., fluorescently labeled ribosomes and tRNAs, modified species-specifically for our new experiments, to study the effect of ABC-Fs on protein synthesis in the presence and absence of different antibiotics (Year 2).The project will shed light on the ABC-F resistance mechanism and how ABC-Fs are regulated.

We will understand why different classes of ABC-Fs have different species specificity, and how the presence of the genes and the gene products affect cell fitness.

The results will be important for rational development of new drugs or procedures to counteract the effect of ABC-Fs, e.g., through combinations with inhibitors. The study will also explicate how transfer of ABC-F genes between bacterial species affects resistance development.