This proposal investigates cell envelope stress responses in Bacillus subtilis, a model organism for the Gram positive bacteria. Antibiotics and other agents that impair the synthesis or function of the cell membrane or peptidoglycan cell wall are of critical importance in clinical medicine. Exposure to sub-lethal levels of antibiotics activates the transcription of sets of genes (cell envelope stress responses) that serve to remodel the cell wall and membrane. In B. subtilis, these stress responses are coordinated by alternative sigma subunits for RNA polymerase of the extracytoplasmic function (ECF) subfamily. This proposal is focused on defining the role of ECF sigma factors in controlling adaptive changes in cell envelope function. One focus will be on the roles of induced lipid biosynthesis pathways and proteins in the genesis and function of lipid rafts. A second focus will be to discover the roles of a newly defined second messenger, cyclic-di-AMP, that is required for cell envelope homeostasis. Using a combination of classical and molecular genetics, genomics, transcriptomics, and biochemical approaches, these studies will provide insights into cell envelope synthesis and function, and reveal novel mechanisms of antibiotic resistance.

Public Health Relevance

The bacterial envelope, comprising a lipid membrane and a peptidoglycan cell wall, separates the cell from the environment and is a target for the majority of antibiotics currently in clinical use. This proposal identifies pathways by which bacteria respond to antibiotic threats by modifying the biosynthesis of the cell envelope with implications for the development of antibiotic resistance. This proposal focuses specifically on the model Gram positive bacterium Bacillus subtilis which is related to Staphylococcus aureus and pathogenic streptococcal and enterococcal strains.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Prokaryotic Cell and Molecular Biology Study Section (PCMB)
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Reddy, Michael K
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Cornell University
Schools of Earth Sciences/Natur
United States
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Lee, Yong Heon; Helmann, John D (2014) Mutations in the primary sigma factor ?A and termination factor rho that reduce susceptibility to cell wall antibiotics. J Bacteriol 196:3700-11
Kingston, Anthony W; Zhao, Heng; Cook, Gregory M et al. (2014) Accumulation of heptaprenyl diphosphate sensitizes Bacillus subtilis to bacitracin: implications for the mechanism of resistance mediated by the BceAB transporter. Mol Microbiol 93:37-49
Kashyap, Des Raj; Rompca, Annemarie; Gaballa, Ahmed et al. (2014) Peptidoglycan recognition proteins kill bacteria by inducing oxidative, thiol, and metal stress. PLoS Pathog 10:e1004280
Lee, Yong Heon; Nam, Ki Hyun; Helmann, John D (2013) A mutation of the RNA polymerase ?' subunit (rpoC) confers cephalosporin resistance in Bacillus subtilis. Antimicrob Agents Chemother 57:56-65
Ringus, Daina L; Gaballa, Ahmed; Helmann, John D et al. (2013) Fluoro-phenyl-styrene-sulfonamide, a novel inhibitor of ýýB activity, prevents the activation of ýýB by environmental and energy stresses in Bacillus subtilis. J Bacteriol 195:2509-17
Lee, Yong Heon; Kingston, Anthony W; Helmann, John D (2012) Glutamate dehydrogenase affects resistance to cell wall antibiotics in Bacillus subtilis. J Bacteriol 194:993-1001
Luo, Yun; Helmann, John D (2012) Analysis of the role of Bacillus subtilis ?(M) in ?-lactam resistance reveals an essential role for c-di-AMP in peptidoglycan homeostasis. Mol Microbiol 83:623-39
Salzberg, Letal I; Luo, Yun; Hachmann, Anna-Barbara et al. (2011) The Bacillus subtilis GntR family repressor YtrA responds to cell wall antibiotics. J Bacteriol 193:5793-801
Sevim, Elif; Gaballa, Ahmed; Belduz, A Osman et al. (2011) DNA-binding properties of the Bacillus subtilis and Aeribacillus pallidus AC6 ýý(D) proteins. J Bacteriol 193:575-9
Guariglia-Oropeza, Veronica; Helmann, John D (2011) Bacillus subtilis ýý(V) confers lysozyme resistance by activation of two cell wall modification pathways, peptidoglycan O-acetylation and D-alanylation of teichoic acids. J Bacteriol 193:6223-32

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