Project Summary Metal dysbiosis is detrimental to bacterial survival as approximately 40% of proteins use metals as a cofactor or structural component, yet metals are often overlooked external stimuli. Due to metal binding being promiscuous and proteins having specific metallo-requirements, bacteria have evolved import and export systems to maintain the homeostasis of biologically relevant metals, with zinc and especially copper tending to be more toxic in moderate concentrations. As metal import and export are dynamic processes with fundamental gaps in knowledge, a multi-omics approach will investigate the pathways for bacterial metal homeostasis for copper and zinc at varying concentrations. The research objectives are to 1) determine how metal influx, efflux, and internal concentrations (the “metallome”) respective to copper and zinc affect the transcriptome and metabolome; 2) build a web-based platform to sort through this data efficiently, and 3) use the data to support and form new hypotheses that examine the interplay of copper and zinc within bacterial systems. These data will also underpin foundational projects for undergraduates in the National Summer Undergraduate Research Project (NSURP), a virtual summer research program for underrepresented minorities that the PI established in 2020. This supplement will also be used to support a program director for NSURP to solidify the foundation further and expand upon its goals.