Posttranslational modification is a rapid and efficient way in which cells adapt to changes in their environment. Biologists at large are interested in advancing our understanding of the mechanisms through which chemical modifications exert their effects, and want to learn more about the impact that these modifications have on the dynamics of the complex metabolic network of the cell. The long-term goal of the work supported by GM62203 is to understand the contributions of the sirtuin- dependent protein acylation/deacylation system (SDPADS) to prokaryotic cell physiology. Previous NIH funding of this project allowed us to advance our understanding of the role of the SDPADS in Gram- negative and Gram-positive bacteria. We are now in an excellent position to perform structure-function analyses of the acetyltransferase of the SDPADS, to dissect the regulation of expression of the genes encoding SDPADS functions, to identify the function of previously unknown genes also involved in acetylation control of protein activity, to establish a role for N-Lys protein acetylation/deacetylation in the control of central metabolic pathways, and to determine if prokaryotes use the SDPADS to directly control gene expression by modulating the activity of transcription regulators. We will take a multidisciplinary approach to answer fundamental questions of prokaryotic cell physiology. The first two aims of this proposal focus on newly discovered effects of acetylation on metabolism and gene expression, and the last two aims focus on the biochemical and structural characterization of the SDPADS enzymes and on the regulation of expression of the genes encoding SDPADS functions. Given the ubiquity of the SDPADS, advances in this research area will have broad impact on our understanding of the physiology of all cells.

Public Health Relevance

. Rapid control of biological activities of proteins is critical for the survival of cells in a changing environment. Posttranslational modifications of lysine residues have a profound effect on gene expression and metabolism. The proposed work will advance our understanding of the mechanism of function of the modifying/demodifying enzymes that comprise a system conserved in all forms of life. Regulation of expression of the genes encoding the modifying/demodifying enzymes is part of the proposed studies. In this project we seek to validate putative new protein substrates of the system, including DNA-binding regulatory proteins.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
7R01GM062203-12
Application #
8320330
Study Section
Special Emphasis Panel (ZRG1-IDM-S (02))
Program Officer
Gerratana, Barbara
Project Start
2001-01-01
Project End
2014-08-31
Budget Start
2012-09-01
Budget End
2014-08-31
Support Year
12
Fiscal Year
2012
Total Cost
$320,199
Indirect Cost
$104,577
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
Mitchell, Carter A; Tucker, Alex C; Escalante-Semerena, Jorge C et al. (2015) The structure of S. lividans acetoacetyl-CoA synthetase shows a novel interaction between the C-terminal extension and the N-terminal domain. Proteins 83:575-81
Crosby, Heidi A; Escalante-Semerena, Jorge C (2014) The acetylation motif in AMP-forming Acyl coenzyme A synthetases contains residues critical for acetylation and recognition by the protein acetyltransferase pat of Rhodopseudomonas palustris. J Bacteriol 196:1496-504
Tucker, Alex C; Escalante-Semerena, Jorge C (2014) Determinants within the C-terminal domain of Streptomyces lividans acetyl-CoA synthetase that block acetylation of its active site lysine in vitro by the protein acetyltransferase (Pat) enzyme. PLoS One 9:e99817
You, Di; Yao, Li-Li; Huang, Dan et al. (2014) Acetyl coenzyme A synthetase is acetylated on multiple lysine residues by a protein acetyltransferase with a single Gcn5-type N-acetyltransferase (GNAT) domain in Saccharopolyspora erythraea. J Bacteriol 196:3169-78
Tucker, Alex C; Escalante-Semerena, Jorge C (2013) Acetoacetyl-CoA synthetase activity is controlled by a protein acetyltransferase with unique domain organization in Streptomyces lividans. Mol Microbiol 87:152-67
Stuecker, Tara N; Hodge, Kelsey M; Escalante-Semerena, Jorge C (2012) The missing link in coenzyme A biosynthesis: PanM (formerly YhhK), a yeast GCN5 acetyltransferase homologue triggers aspartate decarboxylase (PanD) maturation in Salmonella enterica. Mol Microbiol 84:608-19
Crosby, Heidi A; Pelletier, Dale A; Hurst, Gregory B et al. (2012) System-wide studies of N-lysine acetylation in Rhodopseudomonas palustris reveal substrate specificity of protein acetyltransferases. J Biol Chem 287:15590-601
Chan, Chi Ho; Garrity, Jane; Crosby, Heidi A et al. (2011) In Salmonella enterica, the sirtuin-dependent protein acylation/deacylation system (SDPADS) maintains energy homeostasis during growth on low concentrations of acetate. Mol Microbiol 80:168-83
Thao, Sandy; Escalante-Semerena, Jorge C (2011) Biochemical and thermodynamic analyses of Salmonella enterica Pat, a multidomain, multimeric N(?)-lysine acetyltransferase involved in carbon and energy metabolism. MBio 2:
Thao, Sandy; Escalante-Semerena, Jorge C (2011) Control of protein function by reversible Nýý-lysine acetylation in bacteria. Curr Opin Microbiol 14:200-4

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