The goal of this research is to discern the mechanisms by which physiological signals in the environment within the human host are converted to molecular interactions that govern the expression of virulence genes. The model system to be analyzed is the Vibrio cholerae ToxR virulence regulon, for which a number of parameters that influence gene expression, as well as many of the regulators and target genes, are known and partially characterized. The target virulence genes include the tcp operon, toxT, and other """"""""ToxR activated"""""""" genes present on the Vibrio cholerae pathogenicity island, as well as the ctx operon present on a lysogenic bacteriophage. The regulators are encoded by genes distributed around the genome, including toxRS, aphA, aphB, hns, and crp, as well as the tcpPH and toxT genes present on the pathogenicity island. It has recently been determined that multiple regulators function at each target gene promoter. The current proposal focuses on a subset of target promoters and regulators to understand how these regulators function in an interaction with growth condition signals, the promoters, and with each other to control gene expression. In addition, the details of a newly discovered feedback pathway that links pilus biogenesis to virulence gene expression will be investigated. This pathway negatively impacts virulence gene expression when pilus formation is disrupted in certain ways. It may represent a new signal for release from the intestine. Correlating virulence gene expression together with regulatory responses that modulate bacterial physiology represents a new approach that utilizes knowledge of the genome to further our understanding of the basis of virulence gene regulation. These experiments will likely reveal novel virulence factor genes that may prove to be useful vaccine or therapeutic targets. A further understanding of virulence gene expression will also help in the development of ways to modulate it either in vivo or in vitro for improved vaccine production or overproduction of virulence factors for structural analyses. These studies will also help us further understand the mechanism of action of small molecule virulence inhibitors, such as virstatin.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI039654-14
Application #
7777799
Study Section
Special Emphasis Panel (ZRG1-IDM-H (02))
Program Officer
Hall, Robert H
Project Start
1996-06-01
Project End
2012-03-31
Budget Start
2010-04-01
Budget End
2011-03-31
Support Year
14
Fiscal Year
2010
Total Cost
$349,410
Indirect Cost
Name
Dartmouth College
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
041027822
City
Hanover
State
NH
Country
United States
Zip Code
03755
Privett, Britney R; Pellegrini, Maria; Kovacikova, Gabriela et al. (2017) Identification of a Small Molecule Activator for AphB, a LysR-Type Virulence Transcriptional Regulator in Vibrio cholerae. Biochemistry 56:3840-3849
Midgett, Charles R; Almagro-Moreno, Salvador; Pellegrini, Maria et al. (2017) Bile salts and alkaline pH reciprocally modulate the interaction between the periplasmic domains of Vibrio cholerae ToxR and ToxS. Mol Microbiol 105:258-272
Kovacikova, Gabriela; Lin, Wei; Taylor, Ronald K et al. (2017) The Fatty Acid Regulator FadR Influences the Expression of the Virulence Cascade in the El Tor Biotype of Vibrio cholerae by Modulating the Levels of ToxT via Two Different Mechanisms. J Bacteriol 199:
Almagro-Moreno, Salvador; Kim, Tae K; Skorupski, Karen et al. (2015) Proteolysis of virulence regulator ToxR is associated with entry of Vibrio cholerae into a dormant state. PLoS Genet 11:e1005145
Almagro-Moreno, Salvador; Pruss, Kali; Taylor, Ronald K (2015) Intestinal Colonization Dynamics of Vibrio cholerae. PLoS Pathog 11:e1004787
Almagro-Moreno, Salvador; Root, Michael Z; Taylor, Ronald K (2015) Role of ToxS in the proteolytic cascade of virulence regulator ToxR in Vibrio cholerae. Mol Microbiol 98:963-76
Shi, Wei; Kovacikova, Gabriela; Lin, Wei et al. (2015) The 40-residue insertion in Vibrio cholerae FadR facilitates binding of an additional fatty acyl-CoA ligand. Nat Commun 6:6032
Almagro-Moreno, Salvador; Taylor, Ronald K (2013) Cholera: Environmental Reservoirs and Impact on Disease Transmission. Microbiol Spectr 1:
Cerda-Maira, Francisca A; Kovacikova, Gabriela; Jude, Brooke A et al. (2013) Characterization of BreR interaction with the bile response promoters breAB and breR in Vibrio cholerae. J Bacteriol 195:307-17
Taylor, Jennifer L; De Silva, Rukman S; Kovacikova, Gabriela et al. (2012) The crystal structure of AphB, a virulence gene activator from Vibrio cholerae, reveals residues that influence its response to oxygen and pH. Mol Microbiol 83:457-70

Showing the most recent 10 out of 38 publications