Brucella melitensis is a human and animal bacterial pathogen of global significance. Although bacterial factors mediating the host-pathogen interaction have been revealed, host factors that are important for bacterial infection of mammalian cells remain obscure. We have recently exploited a novel Drosophila melanogaster S2 cell model of Brucella infection to perform a pilot screen for host factors that support the replication of this intracellular pathogen. We uncovered genes that have already been established as being important for Brucella infection of mammalian cells. In addition, we uncovered several novel hits that were validated in mammalian cell models. Here, we propose to exploit these findings to perform a large-scale RNAi screen for additional host factors. Specifically, we aim: (1) To perform a large-scale screen for RNAis that disrupt or enhance B. melitensis infection of Drosophila S2 cells;(2) To classify hits into phenotypic and functional categories, and to compare the results obtained to those from the recently completed Mycobacterium fortuitum and Listeria monocytogenes screens. Finally, we shall employ siRNA technology to examine whether the mammalian orthologs of the hits obtained in our Drosophila S2 cell screen mediate Brucella infection of mammalian cells. Taken together, these experiments will define additional host factors that support the uptake and replication of B. melitensis into animal cells, and thereby provide new insights into the molecular mechanisms mediating this important host-pathogen interaction.

Public Health Relevance

This proposal shall identify and characterize novel host factors mediating Brucella infection of animal cells. We expect that this effort shall benefit public health by contributing significantly to our understanding of this pathogen of global significance, and by discovering potential protein targets for therapeutic intervention.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21AI072446-02
Application #
7568748
Study Section
Special Emphasis Panel (ZRG1-IDM-A (90))
Program Officer
Mukhopadhyay, Suman
Project Start
2008-02-15
Project End
2012-01-31
Budget Start
2009-02-01
Budget End
2012-01-31
Support Year
2
Fiscal Year
2009
Total Cost
$180,232
Indirect Cost
Name
Texas A&M University
Department
Other Basic Sciences
Type
Schools of Earth Sciences/Natur
DUNS #
078592789
City
College Station
State
TX
Country
United States
Zip Code
77845
Pandey, Aseem; Lin, Furong; Cabello, Ana L et al. (2018) Activation of Host IRE1?-Dependent Signaling Axis Contributes the Intracellular Parasitism of Brucella melitensis. Front Cell Infect Microbiol 8:103
Criscitiello, Michael F; Dickman, Martin B; Samuel, James E et al. (2013) Tripping on acid: trans-kingdom perspectives on biological acids in immunity and pathogenesis. PLoS Pathog 9:e1003402
Criscitiello, Michael F; de Figueiredo, Paul (2013) Fifty shades of immune defense. PLoS Pathog 9:e1003110
Jupiter, Daniel C; Ficht, Thomas A; Samuel, James et al. (2010) DNA watermarking of infectious agents: progress and prospects. PLoS Pathog 6:e1000950