This project will analyze the function of the LcrE protein in Yersinia pestis. LcrE is implicated in the regulation of a unique secretion mechanism for virulence proteins, the Yop proteins of Yersinia. The Yops lack identifiable signal sequences and are secreted without proteolytic processing. Expression and secretion of Yops in vitro regulated by calcium. Eucaryotic cells triggers the translocation of at least one Yop directly from the surface attached bacterium into the host cell. This appears to be a mechanism of broad interest since it appears to be present in other pathogens as well. This application proposes to identify functional domains of LcrE required to target LcrE to the Yop secretion apparatus, identify domains required to regulate Yop secretion, and identify proteins that interact with LcrE. This will be accomplished by deletion analysis of LcrE, characterization of second- site mutations to identify proteins that interact with LcrE. Protein affinity chromatography, affinity blotting and immunoprecipitation will be used to confirm these interactions. Finally, the effect of LcrE mutants on the translocation of YopE into eucaryotic cells will be determined. These experiments are designed to map regions and residues of LcrE to specific functions and reveal how LcrE functions.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29AI039575-05
Application #
6169699
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Program Officer
Baker, Phillip J
Project Start
1996-06-01
Project End
2001-05-31
Budget Start
2000-06-01
Budget End
2001-05-31
Support Year
5
Fiscal Year
2000
Total Cost
$111,991
Indirect Cost
Name
University of Miami School of Medicine
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
City
Miami
State
FL
Country
United States
Zip Code
33146
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Minato, Yusuke; Ghosh, Amit; Faulkner, Wyatt J et al. (2013) Na+/H+ antiport is essential for Yersinia pestis virulence. Infect Immun 81:3163-72
Chaudhury, Sukanya; Battaile, Kevin P; Lovell, Scott et al. (2013) Structure of the Yersinia pestis tip protein LcrV refined to 1.65?Å resolution. Acta Crystallogr Sect F Struct Biol Cryst Commun 69:477-81
Joseph, Sabrina S; Plano, Gregory V (2013) The SycN/YscB chaperone-binding domain of YopN is required for the calcium-dependent regulation of Yop secretion by Yersinia pestis. Front Cell Infect Microbiol 3:1
Bartra, Sara Schesser; Gong, Xin; Lorica, Cherish D et al. (2012) The outer membrane protein A (OmpA) of Yersinia pestis promotes intracellular survival and virulence in mice. Microb Pathog 52:41-6