The long-term goal of our research is to dissect the mechanism of iron uptake and the regulation of its expression in the pathogens Vibrio anguillarum and V. vulnificus. In the fish pathogen V. anguillarum the pJM1 mediated iron uptake system plays an essential role in systemic infection. This system is an excellent model due to its striking similarities to human septicemic diseases, such as those caused by V. vulnificus in iron-overloaded patients. Even under these high iron conditions, there are iron-limiting niches at which iron limitation-induced genes play a vital role in virulence. In this competitive renewal we will complete the analysis of the regulation of the plasmid-mediated iron uptake genes of V. anguillarum and dissect the mechanism of iron transport in V. anguillarum and V. vulnificus. We have previously characterized the essential components of the V. anguillarum iron uptake system: the siderophore anguibactin and the proteins and regulatory elements controlling its synthesis and transport. We also characterized iron-regulated genes crucial to the virulence of V. vulnificus. We propose: 1. Mechanism of regulation of the ITB operon by AngR and RNAb. AngR is a bifunctional protein that intervenes in anguibactin biosynthesis and acts in the regulation of expression of the iron transport-biosynthesis (ITB) operon. We intend to: a) complete the analysis of the regulatory domains of AngR and b) characterize the structure-function of RNAb in the regulating transcription within the ITB operon. 2. Molecular characterization of the mechanisms of TonB2- mediated iron transport and virulence in V. anguillarum. Of the two tonB systems available to transduce the energy of the proton motive force of the cytoplasmic membrane of V. anguillarum, only the tonB2 system is required for virulence. Since we recently solved the structure of the TonB2 protein, we intend to identify residues important for the TonB2 recognition of the FatA outer membrane receptor and to identify how specific mutations in FatA affect virulence. 3. Characterization of the TtpC function in the TonB2-mediated iron transport and virulence in V. anguillarum and V. vulnificus. We identified a gene upstream of tonB2 in V. anguillarum, V. vulnificus, and V. cholerae that encodes a novel protein essential for tonB2-mediated iron transport. We will characterize this protein, TtpC, to understand its role in iron transport and virulence of V. anguillarum and V. vulnificus.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI019018-28
Application #
8089586
Study Section
Special Emphasis Panel (ZRG1-IDM-A (02))
Program Officer
Hall, Robert H
Project Start
1982-04-01
Project End
2013-06-30
Budget Start
2011-07-01
Budget End
2013-06-30
Support Year
28
Fiscal Year
2011
Total Cost
$370,170
Indirect Cost
Name
Oregon Health and Science University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
096997515
City
Portland
State
OR
Country
United States
Zip Code
97239
Naka, Hiroaki; Liu, Moqing; Actis, Luis A et al. (2013) Plasmid- and chromosome-encoded siderophore anguibactin systems found in marine vibrios: biosynthesis, transport and evolution. Biometals 26:537-47
Naka, Hiroaki; Liu, Moqing; Crosa, Jorge H (2013) Two ABC transporter systems participate in siderophore transport in the marine pathogen Vibrio anguillarum 775 (pJM1). FEMS Microbiol Lett 341:79-86
Naka, Hiroaki; Chen, Qian; Mitoma, Yasutami et al. (2012) Two replication regions in the pJM1 virulence plasmid of the marine pathogen Vibrio anguillarum. Plasmid 67:95-101
Naka, Hiroaki; Crosa, Jorge H (2012) Identification and characterization of a novel outer membrane protein receptor FetA for ferric enterobactin transport in Vibrio anguillarum 775 (pJM1). Biometals 25:125-33
Kustusch, Ryan J; Kuehl, Carole J; Crosa, Jorge H (2012) The ttpC gene is contained in two of three TonB systems in the human pathogen Vibrio vulnificus, but only one is active in iron transport and virulence. J Bacteriol 194:3250-9
Di Lorenzo, Manuela; Stork, Michiel; Crosa, Jorge H (2011) Genetic and biochemical analyses of chromosome and plasmid gene homologues encoding ICL and ArCP domains in Vibrio anguillarum strain 775. Biometals 24:629-43
Thompson, Fabiano L; Thompson, Cristiane C; Dias, Graciela M et al. (2011) The genus Listonella MacDonell and Colwell 1986 is a later heterotypic synonym of the genus Vibrio Pacini 1854 (Approved Lists 1980)--a taxonomic opinion. Int J Syst Evol Microbiol 61:3023-7
Naka, Hiroaki; Crosa, Jorge H (2011) Genetic Determinants of Virulence in the Marine Fish Pathogen Vibrio anguillarum. Fish Pathol 46:1-10
Kustusch, Ryan J; Kuehl, Carole J; Crosa, Jorge H (2011) Power plays: iron transport and energy transduction in pathogenic vibrios. Biometals 24:559-66
Naka, Hiroaki; Dias, Graciela M; Thompson, Cristiane C et al. (2011) Complete genome sequence of the marine fish pathogen Vibrio anguillarum harboring the pJM1 virulence plasmid and genomic comparison with other virulent strains of V. anguillarum and V. ordalii. Infect Immun 79:2889-900

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