Aeromonas is an emerging human pathogen, which causes gastroenteritis and septicemia. The organism is being isolated in increasing numbers from food and water, and is becoming resistant to chlorination in water and to multiple antibiotics. The focus of this grant is on a cytotoxic enterotoxin (Act)of Aeromonas, which, in addition to causing gastroenteritis, leads to fatal, non-intestinal infections. Based on the data generated during the current funding period, the following specific aims will be addressed.
In Aim 1, we will identify a protein/glycoprotein receptor, which appears to be attached to the plasma membrane by a GPI-anchored protein on the intestinal epithelial cell line (T84), to which Act binds, to initiate a signal transduction cascade. These studies will be accomplished by photoaffinity labeling the receptor with Act, by a genetic approach based on a yeast two-hybrid system, and/or by surface plasmon resonance.
In Aim 2, we will delineate the signal transduction cascade triggered by binding of Act to its cell surface receptor on the host cell to better understand the mechanism of action of Act. These studies will involve examining the effect of calcium mobilization and oxidative-stress pathways on TNFalpha and PGE2 production in Act-stimulated cells.
In Aim 3, the intracellular trafficking of Act in the host cell will be examined by using specific inhibitors and the mutated Rab proteins using immunofluorescence/confocal microscopy, and we will study stress-associated protein kinase activation by Act from within the host cell. For these studies, Act will be delivered into T84 cells via lipofection or electroporation to prevent receptor-mediated signaling.
In Aim 4, we will dissect the role of various mediators generated via Act signaling that lead to different biological effects of Act. We will specifically examine mechanism of Act-induced apoptosis in macrophages via caspase 9 and the role of various biological mediators in fluid secretion, using both in vitro and in vivo models. These studies will provide valuable information in intervening in the severe pathological sequelae associated with Aeromonas infections in the future.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI041611-08
Application #
7013978
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Program Officer
Hall, Robert H
Project Start
1997-07-01
Project End
2008-02-28
Budget Start
2006-03-01
Budget End
2008-02-28
Support Year
8
Fiscal Year
2006
Total Cost
$221,177
Indirect Cost
Name
University of Texas Medical Br Galveston
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
800771149
City
Galveston
State
TX
Country
United States
Zip Code
77555
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Kozlova, Elena V; Khajanchi, Bijay K; Sha, Jian et al. (2011) Quorum sensing and c-di-GMP-dependent alterations in gene transcripts and virulence-associated phenotypes in a clinical isolate of Aeromonas hydrophila. Microb Pathog 50:213-23
Sierra, Johanna C; Suarez, Giovanni; Sha, Jian et al. (2010) Unraveling the mechanism of action of a new type III secretion system effector AexU from Aeromonas hydrophila. Microb Pathog 49:122-34
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Suarez, G; Sierra, J C; Erova, T E et al. (2010) A type VI secretion system effector protein, VgrG1, from Aeromonas hydrophila that induces host cell toxicity by ADP ribosylation of actin. J Bacteriol 192:155-68

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