Many thanks to many members of the Yang Lab who contributed to this project (including many undergrads!) and to our collaborators Douglas McCloskey and Xiaoyang Su! 11/11

@njms-mdphd.bsky.social

We're eager to explore these topics in future projects and to see how generalizable these phenotypes are to pathogenic E coli and other species. We think these mechanisms may inform design of antibiotic adjuvants that can slow or prevent resistance development. 10/n

Importantly, little is known on how energy balance alters bacterial physiology. Although metabolic stress is known to inhibit growth and enhance metabolism, our work is the first to study stress responses and antibiotic phenotypes caused by disrupted energy balance. 9/n

By developing a new assay, Barry showed that mismatches between ATP utilization and production (disrupting ATP homeostasis) are sufficient for augmenting antibiotic killing. Thus, Barry's work introduces new concepts in understanding the interplay between energy balance and antibiotic efficacy. 8/n

Using bacterial genetics approaches, Barry was able to show that bioenergetic stress mechanistically induces stress responses that increase antibiotic stress-induced mutation rates and that protect against antibiotic killing. 7/n

This was also surprising that we previously showed that high levels of reactive oxygen species (ROS) were associated with high antibiotic killing, but the pF1 and pNOX cells had decreased killing while also possessing high levels of ROS! 6/n

Correlations between ATP availability and persistence are known and are usually thought about as correlations between metabolic activity (e.g., dormancy, low ATP) and killing efficacy. But Barry's cells have HIGH metabolic activity, which we previously found to INCREASE antibiotic killing. 5/n

Surprisingly, Barry found that bioenergetically stressed cells evolve antibiotic resistance FASTER than control cells (!!) and are also in general protected from bactericidal antibiotic stress (persistence). 4/n

These strains constitutively hydrolyze ATP (pF1) or NADH (pNOX), creating ATP or NADH sinks in these cells. These in turn induce 'bioenergetic stress' in which the cells are always struggling to meet their energetic demands. 3/n

Barry generated a genetic model system (frequently used by the metabolic engineering community) involving E coli cells over-expressing ATP synthase F1 complex genes (pF1) or heterologously expressing NADH oxidase from Streptococcus (pNOX). 2/n

Would you mind adding me too please? Thanks in advance!