Measuring Escherichia coli Gene Expression during Human Urinary Tract Infections

Abstract

Extraintestinal Escherichia coli (E. coli) evolved by acquisition of pathogenicity islands, phage, plasmids, and DNA segments by horizontal gene transfer. Strains are heterogeneous but virulent uropathogenic isolates more often have specific fimbriae, toxins, and iron receptors than commensal strains. One may ask whether it is the virulence factors alone that are required to establish infection. While these virulence factors clearly contribute strongly to pathogenesis, bacteria must survive by metabolizing nutrients available to them. By constructing mutants in all major metabolic pathways and co-challenging mice transurethrally with each mutant and the wild type strain, we identified which major metabolic pathways are required to infect the urinary tract. We must also ask what else is E. coli doing in vivo? To answer this question, we examined the transcriptome of E. coli CFT073 in the murine model of urinary tract infection (UTI) as well as for E. coli strains collected and analyzed directly from the urine of patients attending either a urology clinic or a university health clinic for symptoms of UTI. Using microarrays and RNA-seq, we measured in vivo gene expression for these uropathogenic E. coli strains, identifying genes upregulated during murine and human UTI. Our findings allow us to propose a new definition of bacterial virulence.

Keywords: E. coli; RNA-seq; in vivo gene expression; urinary tract infection.

Figure 1

Imaging of uropathogenic E. coli during urinary tract infection (UTI).

Figure 2

Prevalence of virulence genes found on pathogenicity islands.

Figure 3

Virulence gene expression by E. coli in urine of patients with UTI.

Figure 4

E. coli virulence gene transcript levels during human UTI. (A) Results from E. coli HM27; (B) Results from E. coli HM69.

Figure 4

E. coli virulence gene transcript levels during human UTI. (A) Results from E. coli HM27; (B) Results from E. coli HM69.