Analysis of Metabolic Stress and Functional Integration in Prokaryotes

PROJECT SUMMARY A fundamental feature of a living system is its integrated network of biochemical pathways that respond to endogenous and environmental stresses. In humans, there is a strong connection between metabolic network dysfunction and disease, while in microbes metabolic network structure dictates organismal capabilities, including pathogenesis. Metabolic strategies are conserved across biology, and insights obtained from model organisms provide the means to advance our understanding of general metabolic paradigms, which can often be extrapolated to higher organisms including humans. The PI's laboratory utilizes a broad approach that includes in vivo genetics, molecular biology, biochemistry, bioinformatics, proteomics, and metabolomics and other global approaches to address timely questions in metabolism. In the long-term, the PI's research will contribute to the understanding of the structure and integration of bacterial metabolic networks, how they respond to perturbation, and how specialized capabilities (e.g., causing disease) are integrated into metabolism. Reaching this vision will require the rigorous definition of metabolic components and their integration, which together form the complex system of metabolism. Knowledge of fundamental metabolic processes and a mechanistic understanding of functional components is critical to efforts aimed at treating metabolic diseases, defining pathogenic strategies, and targeting metabolism for rational drug design, synthetic biology, microbiome research, etc. Efforts in the next five years focus on, i) extending the understanding of the RidA stress system, ii) comparing the endogenous stress systems of multiple microbes, iii) identifying functions of additional Rid protein family members, iv) integrating results from these studies into the metabolism of pyridoxal phosphate, and v) defining a function for the conserved YggS protein. Notably, the human YggS homolog (i.e., PROSC) is a biomarker in B6 epilepsy in humans. The goals of this proposal will be accomplished through a combination of chemical, biochemical, molecular genetics, bioinformatics and global approaches. The work here is motivated by our desire to understand the complex system of metabolism, specifically understanding metabolic stress generated by the production of reactive metabolites during growth, how such stress can damage cellular components if not neutralized and discovering a unifying role for the members of the broadly conserved Rid protein superfamily family.