Salmonella enterica represents a considerable burden to human and animal health worldwide. A significant effort has been made to understand the pathogenesis of this intracellular pathogen and the host factors that mediate host defense. Clinical and experimental evidence have unequivocally demonstrated that CD4+ T cells and IFN3 are critical for preventing systemic disease by non-typhoidal Salmonella. IFN3 likely exerts diverse functions in resistance to this intracellular bacterium, including the activation of the antimicrobial arsenal of macrophages. Recent studies have indicated that IFN3 synergizes with Salmonella ligands to enhance the transcription of iNOS. The resultant high NO synthesis mediates most of the profound and long-lasting anti-Salmonella activity of IFN3-primed macrophages. The molecular mechanism(s) by which IFN3- activated NO synthesis enhances the anti-Salmonella activity of macrophages remains, however, largely unknown. We have generated biochemical and genetic evidence in support of a model in which C203 of the SsrB response regulator that mediates global Salmonella pathogenicity island 2 (SPI2) transcription is an important target of reactive nitrogen species (RNS). The goal of this application is to identify the molecular mechanisms underlying the RNS-mediated repression of SPI2 transcription. It is hypothesized that NO congeners repress SPI2 transcription by S-nitrosylating (-SNO) C203 of the dimerization domain of the SsrB response regulator. Specifically, we propose to 1) determine the RNS-mediated modifications that inactivate SsrB regulatory functions;2) examine SPI2 function in the context of Salmonella antinitrosative defenses;3) select for ssrB variant alleles that render SsrB signaling insensitive to RNS;and 4) characterize SsrB residues critical for dimerization.

Public Health Relevance

Salmonella enterica represents a considerable burden to human and animal health worldwide. The mechanisms underlying the pathogenesis of salmonellosis are incompletely understood. The proposed research will shed light on the host pathogen interactions that modulate the expression of a virulence factors key to the ability of Salmonella to cause human disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI054959-10
Application #
8415905
Study Section
Special Emphasis Panel (ZRG1-IDM-S (03))
Program Officer
Alexander, William A
Project Start
2003-09-30
Project End
2014-12-31
Budget Start
2013-01-01
Budget End
2014-12-31
Support Year
10
Fiscal Year
2013
Total Cost
$369,464
Indirect Cost
$127,984
Name
University of Colorado Denver
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
041096314
City
Aurora
State
CO
Country
United States
Zip Code
80045
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