Uropathogenic E. coli (UPEC) are the leading cause of all types of urinary tract infections (UTIs): cystitis and pyelonephritis, community- acquired and nosocomial, complicated and uncomplicated, symptomatic and asymptomatic. Despite many of study, a clear understanding of the pathogenesis of UPEC infections has not emerged. With the exception of type-1 and P fimbriae, heretofore suspected virulence factors have not been confirmed to be essential for UT in animal models. Meanwhile, the discovery the pathogenicity islands in UPEC strains has awakened our appreciation that vast stretches of the genomes of these organisms have yet to be scrutinized. We propose a trifaceted approach to accomplish signature-tagged mutagenesis to identify genetic loci required for colonization in our murine model of ascending UTI. Our success thus far in isolating 12 such mutants portends well for the achievement of this aim. Our second specific aim is to identify fragments of E. coli mutants portends well for the achievement of this aim. Our second specific aim is to identify fragments of E. coli CFT073 genome that are not present in E. coli K-12. Our second specific aim is to identify fragments of E. coli CFT073 genome that are not present in E. coli K-12. We have used a PCR-based genome subtraction approach to clone fragments enriched for such sequences and, using this approach have identified a gene encoding the usher from a previously unknown fimbria. Our third specific aim is to use a green fluorescent protein promoter trap and a fluorescence activated cell sorter to identify promoters from E. coli CFT073 that are expressed specifically in the murine urinary tract. Finally, we will prioritize the loci identified in the first three specific aims and select the most promising for further study in vivo and in vitro to determine the mechanisms by which the products these genes contribute to the pathogenesis of UTI. We believe that this comprehensive analysis of the virulence determinants of UPEC will greatly advance our understanding of the pathogenesis of UTI.

Project Start
2000-09-01
Project End
2001-06-30
Budget Start
Budget End
Support Year
6
Fiscal Year
2000
Total Cost
$129,815
Indirect Cost
Name
University of Maryland Baltimore
Department
Type
DUNS #
003255213
City
Baltimore
State
MD
Country
United States
Zip Code
21201
Buckles, Eric L; Luterbach, Courtney L; Wang, Xiaolin et al. (2015) Signature-tagged mutagenesis and co-infection studies demonstrate the importance of P fimbriae in a murine model of urinary tract infection. Pathog Dis 73:
Buckles, Eric L; Wang, Xiaolin; Lane, M Chelsea et al. (2009) Role of the K2 capsule in Escherichia coli urinary tract infection and serum resistance. J Infect Dis 199:1689-97
Lane, M Chelsea; Li, Xin; Pearson, Melanie M et al. (2009) Oxygen-limiting conditions enrich for fimbriate cells of uropathogenic Proteus mirabilis and Escherichia coli. J Bacteriol 191:1382-92
Zupancic, Margaret L; Frieman, Matthew; Smith, David et al. (2008) Glycan microarray analysis of Candida glabrata adhesin ligand specificity. Mol Microbiol 68:547-59
Jacobsen, Sandra M; Lane, Mary C; Harro, Jean M et al. (2008) The high-affinity phosphate transporter Pst is a virulence factor for Proteus mirabilis during complicated urinary tract infection. FEMS Immunol Med Microbiol 52:180-93
Ma, Biao; Pan, Shih-Jung; Zupancic, Margaret L et al. (2007) Assimilation of NAD(+) precursors in Candida glabrata. Mol Microbiol 66:14-25
Buckles, Eric L; Wang, Xiaolin; Lockatell, C Virginia et al. (2006) PhoU enhances the ability of extraintestinal pathogenic Escherichia coli strain CFT073 to colonize the murine urinary tract. Microbiology 152:153-60
Castano, Irene; Pan, Shih-Jung; Zupancic, Margaret et al. (2005) Telomere length control and transcriptional regulation of subtelomeric adhesins in Candida glabrata. Mol Microbiol 55:1246-58
Domergue, Renee; Castano, Irene; De Las Penas, Alejandro et al. (2005) Nicotinic acid limitation regulates silencing of Candida adhesins during UTI. Science 308:866-70
Jansen, Angela M; Lockatell, Virginia; Johnson, David E et al. (2004) Mannose-resistant Proteus-like fimbriae are produced by most Proteus mirabilis strains infecting the urinary tract, dictate the in vivo localization of bacteria, and contribute to biofilm formation. Infect Immun 72:7294-305

Showing the most recent 10 out of 31 publications