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 #
5R01AI044486-14
Application #
8586515
Study Section
Bacterial Pathogenesis Study Section (BACP)
Program Officer
Alexander, William A
Project Start
1998-12-01
Project End
2016-11-30
Budget Start
2013-12-01
Budget End
2014-11-30
Support Year
14
Fiscal Year
2014
Total Cost
$347,625
Indirect Cost
$122,625
Name
University of Washington
Department
Pathology
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Frawley, Elaine R; Fang, Ferric C (2014) The ins and outs of bacterial iron metabolism. Mol Microbiol 93:609-16
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
Shen, Shu; Fang, Ferric C (2012) Integrated stress responses in Salmonella. Int J Food Microbiol 152:75-81
Casadevall, Arturo; Fang, Ferric C; Pirofski, Liise-Anne (2011) Microbial virulence as an emergent property: consequences and opportunities. PLoS Pathog 7:e1002136
Fang, Ferric C (2011) Antimicrobial actions of reactive oxygen species. MBio 2:
Libby, Stephen J; Brehm, Michael A; Greiner, Dale L et al. (2010) Humanized nonobese diabetic-scid IL2rgammanull mice are susceptible to lethal Salmonella Typhi infection. Proc Natl Acad Sci U S A 107:15589-94
Karlinsey, Joyce E; Maguire, Michael E; Becker, Lynne A et al. (2010) The phage shock protein PspA facilitates divalent metal transport and is required for virulence of Salmonella enterica sv. Typhimurium. Mol Microbiol 78:669-85
Muller, Cecile; Bang, Iel-Soo; Velayudhan, Jyoti et al. (2009) Acid stress activation of the sigma(E) stress response in Salmonella enterica serovar Typhimurium. Mol Microbiol 71:1228-38
Papp-Wallace, Krisztina M; Nartea, Margaret; Kehres, David G et al. (2008) The CorA Mg2+ channel is required for the virulence of Salmonella enterica serovar typhimurium. J Bacteriol 190:6517-23
Fang, Ferric C; Rimsky, Sylvie (2008) New insights into transcriptional regulation by H-NS. Curr Opin Microbiol 11:113-20

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