Knowledge of how most pathogenic microbes adapt to changing conditions within the host is limited. Advances in our understanding of the basic mechanisms of adaptation, gene regulation and virulence gene expression will provide new molecular targets for therapy to reduce the large medical burden imposed by Helicobacter pylori, which chronically colonizes over half of the world's human population and causes gastritis, ulcer disease, gastric carcinoma and mucosa-associated lymphoid tissue (MALT) lymphoma. We have previously shown that H. pylori regulates expression of key virulence factors and a small subset of regulatory proteins in response to environmental stress. We hypothesize that these regulatory factors modulate expression of genes that are crucial for adaptation to environmental stress and are likely to be essential for colonization and/or persistence of H. pyloriwithin the gastric environment. Herein, we propose detailed genetic and biochemical characterization of one of these factors, Fur. Fur is the only one of the regulators affected by both low pH and iron. Additionally, the classic paradigm of Fur regulation established by studies in other bacteria is considerably more complex in H. pylori', Fur regulates gene expression in both its iron-bound and apo forms. Our studies will define and characterize the iron- bound and apo-Fur regulons, and will seek to identify structural determinants of Fur that mediate each of these two modes of regulation. Finally, to expand our knowledge of virulence gene regulation in response to stress, we will identify additional genes involved in expression of the iron- regulated virulence genes cagA and vacA using reporter constructs and a near-saturating transposon library. These studies will fill a fundamentalgap in knowledge concerning theprocess of adaptation and regulation in H. pylori and should provide potential new therapeutic targets for H. pylori. Additionally, they willprovide novelinsight into the unique mechanisms of Fur-regulation utilized by this importantpathogen. Relevance to Public Health: H. pylori infects more than 50% of the worlds population and causes a range of diseases. Our studies will help to shed light on genes involved in the process of surviving in the human body and should provide novel targets for vaccine and therapeutic design.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI065529-05
Application #
7993513
Study Section
Bacterial Pathogenesis Study Section (BACP)
Program Officer
Mills, Melody
Project Start
2006-12-15
Project End
2013-11-30
Budget Start
2010-12-01
Budget End
2013-11-30
Support Year
5
Fiscal Year
2011
Total Cost
$367,285
Indirect Cost
Name
Henry M. Jackson Fdn for the Adv Mil/Med
Department
Type
DUNS #
144676566
City
Bethesda
State
MD
Country
United States
Zip Code
20817
Makobongo, Morris O; Gilbreath, Jeremy J; Merrell, D Scott (2014) Nontraditional therapies to treat Helicobacter pylori infection. J Microbiol 52:259-72
Gilbreath, Jeremy J; Pich, Oscar Q; Benoit, Stéphane L et al. (2013) Random and site-specific mutagenesis of the Helicobacter pylori ferric uptake regulator provides insight into Fur structure-function relationships. Mol Microbiol 89:304-23
Pich, Oscar Q; Merrell, D Scott (2013) The ferric uptake regulator of Helicobacter pylori: a critical player in the battle for iron and colonization of the stomach. Future Microbiol 8:725-38
Carpenter, Beth M; Gilbreath, Jeremy J; Pich, Oscar Q et al. (2013) Identification and characterization of novel Helicobacter pylori apo-fur-regulated target genes. J Bacteriol 195:5526-39
Pich, Oscar Q; Carpenter, Beth M; Gilbreath, Jeremy J et al. (2012) Detailed analysis of Helicobacter pylori Fur-regulated promoters reveals a Fur box core sequence and novel Fur-regulated genes. Mol Microbiol 84:921-41
Whitmire, Jeannette M; Merrell, D Scott (2012) Successful culture techniques for Helicobacter species: establishing H. pylori cultures from infected rodents. Methods Mol Biol 921:29-35
Makobongo, Morris O; Gancz, Hanan; Carpenter, Beth M et al. (2012) The oligo-acyl lysyl antimicrobial peptide C₁₂K-2β₁₂ exhibits a dual mechanism of action and demonstrates strong in vivo efficacy against Helicobacter pylori. Antimicrob Agents Chemother 56:378-90
Gilbreath, Jeremy J; West, Abby L; Pich, Oscar Q et al. (2012) Fur activates expression of the 2-oxoglutarate oxidoreductase genes (oorDABC) in Helicobacter pylori. J Bacteriol 194:6490-7
Whitmire, Jeannette M; Merrell, D Scott (2012) Successful culture techniques for Helicobacter species: general culture techniques for Helicobacter pylori. Methods Mol Biol 921:17-27
Gilbreath, Jeremy J; Colvocoresses Dodds, Jennifer; Rick, Paul D et al. (2012) Enterobacterial common antigen mutants of Salmonella enterica serovar Typhimurium establish a persistent infection and provide protection against subsequent lethal challenge. Infect Immun 80:441-50

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