Pathogenic bacteria must withstand extracytoplasmic stress, or perturbation of the cell envelope and periplasm, to survive in the host environment. An alternative sigma factor called CE (SigmaE, RpoE) controls extracytoplasmic stress responses in Salmonella enterica sv. Typhimurium and other bacteria. We have demonstrated that CE is required for Salmonella virulence and resistance to killing by macrophages and antimicrobial peptides. During the last project period, we have discovered: (a) a novel mechanism of CE activation by acid pH important for survival in macrophages, (b) two new CE-regulated virulence genes encoding a peptidyl-prolyl isomerase and a periplasmic protein of unknown function, and (c) unanticipated interactions linking CE to other stress regulatory networks. The central hypothesis of this proposal is that CE acts in concert with the Rcs, Psp, Cpx and Bae systems to form an integrated regulatory network that defends bacteria from cell envelope damage and maintains pathways of energy generation essential for cellular homeostasis.
The specific aims are to explore the mechanisms underlying our recent findings: 1. Analyze the mechanism by which CE is activated by acid pH. Biochemical and genetic approaches will be used to examine the effects of pH on the conformation of the periplasmic RseA domain and its function in the CE regulatory cascade. 2. Determine the mechanism by which the peptidyl-prolyl isomerase SlpA and the periplasmic protein YggN promote bacterial virulence. Proteomic and phenotypic assays will explore the contributions of SlpA and YggN to Salmonella pathogenesis. 3. Investigate the mechanisms by which the Psp and Rcs systems preserve cell survival under stress conditions. A range of functional and biochemical studies will assess the central role of proton motive force and its preservation by extracytoplasmic stress responses. These studies will provide important new insights into conserved mechanisms of bacterial pathogenesis that have broad biological implications.

Public Health Relevance

Salmonella is an important cause of gastrointestinal and systemic infections in both industrialized and developing countries. This project explores our lab's recent discoveries regarding strategies used by Salmonella to resist external stresses imposed by host defense mechanisms. Characterization of these pathways will provide new insights into bacterial pathogenesis, reveal novel targets that can improve the effectiveness of antibiotics, and lead to more efficacious treatment of bacterial infections.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI044486-12
Application #
8235760
Study Section
Bacterial Pathogenesis Study Section (BACP)
Program Officer
Alexander, William A
Project Start
1998-12-01
Project End
2016-11-30
Budget Start
2011-12-01
Budget End
2012-11-30
Support Year
12
Fiscal Year
2012
Total Cost
$385,521
Indirect Cost
$135,521
Name
University of Washington
Department
Pathology
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Fang, Ferric C; Frawley, Elaine R; Tapscott, Timothy et al. (2016) Bacterial Stress Responses during Host Infection. Cell Host Microbe 20:133-43
Fang, Ferric C; Frawley, Elaine R; Tapscott, Timothy et al. (2016) Discrimination and Integration of Stress Signals by Pathogenic Bacteria. Cell Host Microbe 20:144-53
Kinkel, Traci L; Ramos-Montañez, Smirla; Pando, Jasmine M et al. (2016) An essential role for bacterial nitric oxide synthase in Staphylococcus aureus electron transfer and colonization. Nat Microbiol 2:16224
Singletary, Larissa A; Karlinsey, Joyce E; Libby, Stephen J et al. (2016) Loss of Multicellular Behavior in Epidemic African Nontyphoidal Salmonella enterica Serovar Typhimurium ST313 Strain D23580. MBio 7:e02265
Nairz, Manfred; Schroll, Andrea; Haschka, David et al. (2015) Lipocalin-2 ensures host defense against Salmonella Typhimurium by controlling macrophage iron homeostasis and immune response. Eur J Immunol 45:3073-86
Nairz, Manfred; Ferring-Appel, Dunja; Casarrubea, Daniela et al. (2015) Iron Regulatory Proteins Mediate Host Resistance to Salmonella Infection. Cell Host Microbe 18:254-61
Frawley, Elaine R; Fang, Ferric C (2014) The ins and outs of bacterial iron metabolism. Mol Microbiol 93:609-16
Bäumler, Andreas; Fang, Ferric C (2013) Host specificity of bacterial pathogens. Cold Spring Harb Perspect Med 3:a010041
Moreira, Cristiano G; Herrera, Carmen M; Needham, Brittany D et al. (2013) Virulence and stress-related periplasmic protein (VisP) in bacterial/host associations. Proc Natl Acad Sci U S A 110:1470-5
Frawley, Elaine R; Crouch, Marie-Laure V; Bingham-Ramos, Lacey K et al. (2013) Iron and citrate export by a major facilitator superfamily pump regulates metabolism and stress resistance in Salmonella Typhimurium. Proc Natl Acad Sci U S A 110:12054-9

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