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

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Specialized Center--Cooperative Agreements (U54)
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Special Emphasis Panel (ZAI1-DDS-M (J2))
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University of California Irvine
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Waggoner, Jesse J; Gresh, Lionel; Mohamed-Hadley, Alisha et al. (2016) Single-Reaction Multiplex Reverse Transcription PCR for Detection of Zika, Chikungunya, and Dengue Viruses. Emerg Infect Dis 22:1295-7
Ziegler, Christopher M; Eisenhauer, Philip; Bruce, Emily A et al. (2016) The Lymphocytic Choriomeningitis Virus Matrix Protein PPXY Late Domain Drives the Production of Defective Interfering Particles. PLoS Pathog 12:e1005501
Barbour, Alan G (2016) Infection resistance and tolerance in Peromyscus spp., natural reservoirs of microbes that are virulent for humans. Semin Cell Dev Biol :
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
Levin, Mattias; King, Jasmine J; Glanville, Jacob et al. (2016) Persistence and evolution of allergen-specific IgE repertoires during subcutaneous specific immunotherapy. J Allergy Clin Immunol 137:1535-44
Chomel, Bruno B; Molia, Sophie; Kasten, Rickie W et al. (2016) Isolation of Bartonella henselae and Two New Bartonella Subspecies, Bartonellakoehlerae Subspecies boulouisii subsp. nov. and Bartonella koehlerae Subspecies bothieri subsp. nov. from Free-Ranging Californian Mountain Lions and Bobcats. PLoS One 11:e0148299
Kern, Aurelie; Zhou, Chensheng W; Jia, Feng et al. (2016) Live-vaccinia virus encapsulation in pH-sensitive polymer increases safety of a reservoir-targeted Lyme disease vaccine by targeting gastrointestinal release. Vaccine 34:4507-13
Zeltina, Antra; Bowden, Thomas A; Lee, Benhur (2016) Emerging Paramyxoviruses: Receptor Tropism and Zoonotic Potential. PLoS Pathog 12:e1005390
Waggoner, Jesse J; Ballesteros, Gabriela; Gresh, Lionel et al. (2016) Clinical evaluation of a single-reaction real-time RT-PCR for pan-dengue and chikungunya virus detection. J Clin Virol 78:57-61
Sanman, Laura E; Qian, Yu; Eisele, Nicholas A et al. (2016) Disruption of glycolytic flux is a signal for inflammasome signaling and pyroptotic cell death. Elife 5:e13663

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