Knowledge is very limited with respect to how facultative pathogens, i.e., pathogens that spend portions of their life cycles both inside and outside of the human body, adapt to drastically changing conditions when passing from a human host into an environmental reservoir. Advances in understanding of the basic mechanisms of adaptation during this transition are needed to provide new molecular targets and strategies for limiting transmission, dissemination and environmental persistence, and thus reduce the large medical burden imposed by this diverse group of pathogens. In the facultative, water-borne pathogen Vibrio cholerae (the causative agent of cholera), signaling by the intracellular secondary messenger molecule cyclic diguanylate (c-di-GMP) leads to the repression of virulence genes, and at the same time, the induction of environmental survival genes. At a late stage of infection, just prior to excretion of V. cholerae in watery stool, expression of three genes that encode c-di-GMP synthetases is induced, leading to the hypothesize that c-di-GMP mediates the transition from a state of virulence to one of environmental suitability. In this project the role of c-di- GMP in the biology of V. cholerae late in infection and in the process of dissemination will be determined using genetic and biochemical methods, including some novel methods created for these studies. In addition, the roles of a family of five c-di-GMP-binding proteins in transducing the c-di- GMP signal into biological changes will be determined. It is anticipated that this work will provide a working model of the central regulatory pathway in V. cholerae that represses virulence gene expression and induces expression of genes important for dissemination. It is also anticipated that these studies will have application to a broad range of bacterial pathogens since most contain multiple c-di-GMP synthetic, degradative and sensory proteins.

Public Health Relevance

This project will advance our understanding of the mechanisms bacterial pathogens use to disseminate from infected humans into environmental reservoirs. This information will provide new targets for limiting dissemination of pathogens and for developing environmental control measures.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI045746-13
Application #
8596792
Study Section
Bacterial Pathogenesis Study Section (BACP)
Program Officer
Hall, Robert H
Project Start
2000-03-01
Project End
2016-11-30
Budget Start
2013-12-01
Budget End
2014-11-30
Support Year
13
Fiscal Year
2014
Total Cost
$371,250
Indirect Cost
$146,250
Name
Tufts University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
039318308
City
Boston
State
MA
Country
United States
Zip Code
02111
Dalia, Ankur B; McDonough, EmilyKate; Camilli, Andrew (2014) Multiplex genome editing by natural transformation. Proc Natl Acad Sci U S A 111:8937-42
van Opijnen, Tim; Lazinski, David W; Camilli, Andrew (2014) Genome-Wide Fitness and Genetic Interactions Determined by Tn-seq, a High-Throughput Massively Parallel Sequencing Method for Microorganisms. Curr Protoc Mol Biol 106:7.16.1-7.16.24
Seed, Kimberley D; Lazinski, David W; Calderwood, Stephen B et al. (2013) A bacteriophage encodes its own CRISPR/Cas adaptive response to evade host innate immunity. Nature 494:489-91
Lazinski, David W; Camilli, Andrew (2013) Homopolymer tail-mediated ligation PCR: a streamlined and highly efficient method for DNA cloning and library construction. Biotechniques 54:25-34
Bradley, Evan S; Bodi, Kip; Ismail, Ayman M et al. (2011) A genome-wide approach to discovery of small RNAs involved in regulation of virulence in Vibrio cholerae. PLoS Pathog 7:e1002126
Bishop, Anne L; Camilli, Andrew (2011) Vibrio cholerae: lessons for mucosal vaccine design. Expert Rev Vaccines 10:79-94
Liu, Jane M; Camilli, Andrew (2010) A broadening world of bacterial small RNAs. Curr Opin Microbiol 13:18-23
Pratt, Jason T; Ismail, Ayman M; Camilli, Andrew (2010) PhoB regulates both environmental and virulence gene expression in Vibrio cholerae. Mol Microbiol 77:1595-605
Pratt, Jason T; McDonough, Emilykate; Camilli, Andrew (2009) PhoB regulates motility, biofilms, and cyclic di-GMP in Vibrio cholerae. J Bacteriol 191:6632-42
Nelson, Eric J; Harris, Jason B; Morris Jr, J Glenn et al. (2009) Cholera transmission: the host, pathogen and bacteriophage dynamic. Nat Rev Microbiol 7:693-702

Showing the most recent 10 out of 30 publications