Burkholderia pseudomailei is the causative agent of melioidosis, a global emerging infectious disease, and it is also classified as a category B potential bioterrorism agent. Thousands of cases of melioidosis have occurred in the tropics, and several have been reported in American travelers (civilian and military personnel). Other cases have been documented in Hawaii, and the bacterium has been detected on the Big Island of Hawaii. Using a novel method we recently pioneered to profile global gene-expression in single prokaryotic cells, we analyze the 'transitome' of S. pseudomailei as the bacterium transits through the hostcell during its intracellular infectious lifecycle. Our preliminary data revealed expression of a series of hypothetical proteins and transcriptional regulators in unique infectious stages of B. pseudomailei. Highthrough-put chromosomal mutagenesis of 200 unique hypothetical-protein-encoding genes, expressed in a stage-specific fashion, and initial phenotype screening narrowed the list to 11 hypothetical proteins that decreased virulence and the ability of 6. pseudomailei to infect host-cells. This application proposes to continue this research through two aims: i) To firmly validate the transitomic data by independent methods using reporter-gene-fusion and single-cell real-time RT-PCR and continue with identifying the importance of the 11 hypothetical proteins using host-cell infection models and a murine melioidosis model of infection, and ii) To generate 45 unique regulatory mutants on the B. pseudomailei chromosome to identify their importance in cell and murine melioidosis models. These studies will yield further insight into the functions of unknown virulence genes expressed in a stage-specific fashion in the 6. pseudoma//e/transitome and help elucidate the molecular mechanisms involved in B. pseudomailei intracellular infection.

Public Health Relevance

B. pseudomallei is a potential bioterrorism agent, a risk to homeland security, and a potential burden to public health. The knowledge gained can lead to improved treatment of thousands of melioidosis cases worldwide and, importantly, this will increase biodefense by reducing the potential risk to homeland security and reducing the potential burden to public health. Ultimately, understanding the function of these virulence genes and mechanisms of infection and disease at the molecular level will aid in rational drug and vaccine design

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Specialized Center--Cooperative Agreements (U54)
Project #
5U54AI065359-08
Application #
8458665
Study Section
Special Emphasis Panel (ZAI1-DDS-M (J2))
Project Start
Project End
Budget Start
2012-05-01
Budget End
2013-04-30
Support Year
8
Fiscal Year
2012
Total Cost
$208,477
Indirect Cost
$22,008
Name
University of California Irvine
Department
Type
DUNS #
046705849
City
Irvine
State
CA
Country
United States
Zip Code
92697
Hertz, Tomer; Beatty, P Robert; MacMillen, Zachary et al. (2017) Antibody Epitopes Identified in Critical Regions of Dengue Virus Nonstructural 1 Protein in Mouse Vaccination and Natural Human Infections. J Immunol 198:4025-4035
Marques, Adriana R; Yang, Xiuli; Smith, Alexis A et al. (2017) Citrate Anticoagulant Improves the Sensitivity of Borreliella (Borrelia) burgdorferi Plasma Culture. J Clin Microbiol 55:3297-3299
Parameswaran, Poornima; Wang, Chunling; Trivedi, Surbhi Bharat et al. (2017) Intrahost Selection Pressures Drive Rapid Dengue Virus Microevolution in Acute Human Infections. Cell Host Microbe 22:400-410.e5
Bortell, Nikki; Flynn, Claudia; Conti, Bruno et al. (2017) Osteopontin Impacts West Nile virus Pathogenesis and Resistance by Regulating Inflammasome Components and Cell Death in the Central Nervous System at Early Time Points. Mediators Inflamm 2017:7582437
Barbour, Alan G (2017) Infection resistance and tolerance in Peromyscus spp., natural reservoirs of microbes that are virulent for humans. Semin Cell Dev Biol 61:115-122
Nualnoi, Teerapat; Norris, Michael H; Tuanyok, Apichai et al. (2017) Development of Immunoassays for Burkholderia pseudomallei Typical and Atypical Lipopolysaccharide Strain Typing. Am J Trop Med Hyg 96:358-367
Nakajima, Rie; Escudero, Raquel; Molina, Douglas M et al. (2016) Towards Development of Improved Serodiagnostics for Tularemia by Use of Francisella tularensis Proteome Microarrays. J Clin Microbiol 54:1755-65
Zeltina, Antra; Bowden, Thomas A; Lee, Benhur (2016) Emerging Paramyxoviruses: Receptor Tropism and Zoonotic Potential. PLoS Pathog 12:e1005390
Ziegler, Christopher M; Eisenhauer, Philip; Bruce, Emily A et al. (2016) A novel phosphoserine motif in the LCMV matrix protein Z regulates the release of infectious virus and defective interfering particles. J Gen Virol 97:2084-9
Park, Arnold; Yun, Tatyana; Hill, Terence E et al. (2016) Optimized P2A for reporter gene insertion into Nipah virus results in efficient ribosomal skipping and wild-type lethality. J Gen Virol 97:839-43

Showing the most recent 10 out of 457 publications