A fundamental feature of a living system is an integrated network of biochemical pathways that can respond to stresses applied by the environment. Microbes, particularly those with a genetic system, provide a technically amenable system to characterize metabolic processes and stress responses. Metabolic strategies are conserved across biology, and insights obtained from microbial systems will contribute to our understanding of general metabolic paradigms. The long-term goal of my research is to contribute to the understanding of metabolic components and the processes they participate in. A rigorous understanding of metabolic processes is critical in efforts to predict the response of cells to environmental change, efforts to develop metabolic modeling strategies and efforts targeting metabolism for rational drug design and/or production of small molecules, to name a few. The goal of the work herein is to characterize a metabolic stress that results from reactive metabolites generated during growth, and to understand the family of proteins that neutralize this stress. This study focuses on a bacterial protein (YjgF) that is a member of highly conserved protein family that neutralizes reactive nitrogen species (e.g. enamines). In the current proposal, we will i) identify the stressors neutralized by the YjgF protein, ii) define the molecular consequences of not neutralizing this metabolic stress and iii) explore the relationship between sequence and functional divergence in the YjgF family. The goals of this proposal will be accomplished through the use of modern chemical, biochemical, biophysical, molecular, genetic and bioinformatic techniques. The work proposed here is motivated by our desire to understand the metabolic stress generated during growth by the production of reactive metabolites that can damage cellular components if they are not neutralized.

Public Health Relevance

Metabolism describes the processes required for life in all biological systems. Understanding metabolism is essential for biomedical progress including targeting metabolism for rational drug design and/ or production of small molecules. Our work contributes to this understanding by defining the biochemical function and metabolic role for a family of proteins that is conserved from bacteria to man.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM095837-04
Application #
8637088
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Barski, Oleg
Project Start
2011-09-01
Project End
2015-05-31
Budget Start
2013-06-01
Budget End
2014-05-31
Support Year
4
Fiscal Year
2013
Total Cost
$239,870
Indirect Cost
$75,820
Name
University of Georgia
Department
Microbiology/Immun/Virology
Type
Schools of Arts and Sciences
DUNS #
004315578
City
Athens
State
GA
Country
United States
Zip Code
30602
Ernst, Dustin C; Downs, Diana M (2018) Mmf1p Couples Amino Acid Metabolism to Mitochondrial DNA Maintenance in Saccharomyces cerevisiae. MBio 9:
Irons, Jessica; Hodge-Hanson, Kelsey M; Downs, Diana M (2018) PA5339, a RidA Homolog, Is Required for Full Growth in Pseudomonas aeruginosa. J Bacteriol 200:
Ernst, Dustin C; Borchert, Andrew J; Downs, Diana M (2018) Perturbation of the metabolic network in Salmonella enterica reveals cross-talk between coenzyme A and thiamine pathways. PLoS One 13:e0197703
Hodge-Hanson, Kelsey M; Zoino, Allison; Downs, Diana M (2018) Expression of Pyridoxal 5'-Phosphate-Independent Racemases Can Reduce 2-Aminoacrylate Stress in Salmonella enterica. J Bacteriol 200:
Ernst, Dustin C; Christopherson, Melissa R; Downs, Diana M (2018) Increased Activity of Cystathionine ?-Lyase Suppresses 2-Aminoacrylate Stress in Salmonella enterica. J Bacteriol 200:
Borchert, Andrew J; Downs, Diana M (2017) Endogenously generated 2-aminoacrylate inhibits motility in Salmonella enterica. Sci Rep 7:12971
Borchert, Andrew J; Downs, Diana M (2017) The Response to 2-Aminoacrylate Differs in Escherichia coli and Salmonella enterica, despite Shared Metabolic Components. J Bacteriol 199:
Hodge-Hanson, Kelsey M; Downs, Diana M (2017) Members of the Rid protein family have broad imine deaminase activity and can accelerate the Pseudomonas aeruginosa D-arginine dehydrogenase (DauA) reaction in vitro. PLoS One 12:e0185544
Ernst, Dustin C; Downs, Diana M (2016) 2-Aminoacrylate Stress Induces a Context-Dependent Glycine Requirement in ridA Strains of Salmonella enterica. J Bacteriol 198:536-43
ElRamlawy, Kareem Gamal; Fujimura, Takashi; Baba, Koji et al. (2016) Der f 34, a Novel Major House Dust Mite Allergen Belonging to a Highly Conserved Rid/YjgF/YER057c/UK114 Family of Imine Deaminases. J Biol Chem 291:21607-21615

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