The proposed work on the role of N-Lysine acetylation on protein function has potentially broad implications of interest to all biologists studying central metabolism and regulation of gene expression as it pertains to cancer and human aging. We have an excellent model prokaryotic organism (Salmonella enterica) in which to perform the proposed work. The chief advantage of working with this bacterium is our ability to study the effect of mutations of interest in diverse genetic backgrounds from which we can learn about accessory functions needed for the acetylation/deacetylation system to be efficient. Use of the S. enterica sophisticated genetic system will greatly facilitate the engineering of strains with required functionalities. This model organism is also extremely valuable for the production and isolation of modified or unmodified proteins whose function can be studied in detail in isolation. The connection between protein acetylation and central metabolism has been established in bacteria, murine and human cells. Thus, results from studies in S. enterica will likely lead to important advances in our understanding of metabolic pathway integration in all forms of life. Improved understanding of the bacterial system will in turn facilitate the implementation of physiological strategies for the production of low molecular weight compounds, antibiotics and other secondary metabolites, peptides, enzymes, and proteins of value to the agricultural and pharmaceutical industries, and biotechnology in general. New knowledge obtained from these studies about the function of the enzyme that acetylates proteins as a function of acetyl-CoA levels in the cell will shed light on how enzyme function might be regulated in response to changes in the physiological state of the cell or in response to environmental stimuli that alter CoA homeostasis. This new information will be useful in the design and construction of variant forms of the acetyltransferase with altered response to specific stimuli. These variant proteins may be critical in the genetic engineering of strains with desirable metabolic capabilities.
