The distinct transcriptome of virulence-associated phylogenetic group B2 Escherichia coli.

Escherichia coli strains of phylogenetic group B2 are often associated with urinary tract infections (UTIs) and several other diseases. Recent genomic and transcriptomic analyses have not suggested or identified specific genes required for virulence, but have instead suggested multiple virulence strategies and complex host-pathogen interactions. Previous analyses have not compared core gene expression between phylogenetic groups or between pathogens and nonpathogens within phylogenetic groups. We compared the core gene expression of 35 strains from three phylogenetic groups that included both pathogens and nonpathogens after growth in a medium that allowed comparable growth of both types of strains. K-means clustering suggested a B2 cluster with 17 group B2 strains and two group A strains; an AD cluster with six group A strains, five group D strains and one B2 strain; and four outliers which included the highly studied model uropathogenic E. coli strains UTI89 and CFT073. Half of the core genes were differentially expressed between B2 and AD cluster strains, including transcripts of genes for all aspects of macromolecular synthesis-replication, transcription, translation, and peptidoglycan synthesis-energy metabolism, and environmental-sensing transcriptional regulators. Notably, core gene expression between nonpathogenic and uropathogenic transcriptomes within phylogenetic groups did not differ. If differences between pathogens and nonpathogens exist, then the differences do not require transcriptional reprogramming. In summary, B2 cluster strains have a distinct transcription pattern that involves hundreds of genes. We propose that this transcription pattern is one factor that contributes to virulence. IMPORTANCE Escherichia coli is a diverse species and an opportunistic pathogen that is associated with various diseases, such as urinary tract infections. When examined, phylogenetic group B2 strains are more often associated with these diseases, but the specific properties that contribute to their virulence are not known. From a comparative transcriptomic analysis, we found that group B2 strains grown in a nutrient-rich medium had a distinct transcription pattern, which is the first evidence that core gene expression differs between phylogenetic groups. Understanding the consequences of group B2 transcription pattern will provide important information on basic E. coli biology, the basis for E. coli virulence, and possibly for developing therapies for a majority of urinary tract infections and other group B2-associated diseases.

Facile transduction with P1 phage in Escherichia coli associated with urinary tract infections.

We describe a modification of a standard method that efficiently transduces genes using P1 phage into uropathogenic Escherichia coli strains. This procedure allows utilization of the large KEIO mutant library for analysis of pathogenic E. coli strains.

Genome Sequences of Seven Clade B2 Escherichia coli Strains Associated with Recurrent Urinary Tract Infections in Postmenopausal Women.

We report the genome sequences of seven recently isolated Escherichia coli strains from symptomatic postmenopausal women with a history of recurrent urinary tract infections. We have observed rapid laboratory evolution of strains after isolation. These strains were minimally passaged before analysis to prevent changes during culturing.

Bacterial Growth of Uropathogenic Escherichia coli in Pooled Urine Is Much Higher than Predicted from the Average Growth in Individual Urine Samples.

Urinary tract infections (UTIs), mostly caused by uropathogenic E. coli (UPEC), affect most women, and often recur. Genomic and transcriptomic analyses have not identified a common set of virulence genes, which has suggested complex host-pathogen interactions and multiple virulence mechanisms. One aspect of the host-pathogen interaction is rapid UPEC growth in urine in vivo. When bacterial growth in urine is studied in vitro, urine is pooled, which is assumed to diminish individual variation. We grew one nonpathogenic and two pathogenic E. coli strains in urine from individuals who never had a UTI, had a UTI history but no current infection, and had a UTI history with a current infection. Bacterial growth showed large variations in individual urine samples, and pooled urine often supported significantly more growth than the average growth from individual urine samples. Total nutrient content tended to be higher in current group urine samples than the never and history grouped samples urine. We propose that pooling optimizes a nutrient mixture in the never and history group urine samples, which are often studied, whereas urine from current group individuals may have a more optimal nutrient mixture because of additional nutrient sources. We conclude that a pooled urine is not "an average urine sample," and that the best comparisons of results between labs using pooled urine would also include results with a standardized synthetic urine. IMPORTANCE Urinary tract infections (UTIs) will affect most women, can recur especially in postmenopausal women, and can become antibiotic recalcitrant. Escherichia coli causes most community-acquired UTIs and recurrent UTIs. Current theories of virulence, based on studies of UTI-associated E. coli, propose multiple virulence mechanisms and complex host-pathogen interactions. Studies of bacterial growth in urine samples-one aspect of the host-pathogen interaction-invariably involve pooled urine that are assumed to eliminate variations between individuals. Our results show that a pooled urine is not necessarily an average urine sample, and we suggest that quantitative and qualitative variations in nutrient content are the basis for this discrepancy. Knowledge of growth-promoting urinary components is important for understanding host-pathogen interactions during UTIs and could contribute to developing nonantibiotic-based therapies.

The Nutrient and Energy Pathway Requirements for Surface Motility of Nonpathogenic and Uropathogenic Escherichia coli.

Uropathogenic E. coli (UPEC) is the causative pathogen for most uncomplicated urinary tract infections. Motility is likely to contribute to these infections, and E. coli possesses flagella-dependent swimming motility, flagella-dependent surface motility (often called swarming), and the recently observed pili-dependent surface motility. Surface motility has not been extensively studied, but for the strains that have been tested nonpathogenic E. coli (NPEC) lab strains use pili, NPEC hypermotile derivatives of these lab strains use flagella, and UPEC strains use flagella. Using a representative of these three types of strains, we showed differences in the nutritional and pathway requirements for surface motility with respect to the glucose concentration, the glycolytic pathway utilized, acetogenesis, and the TCA cycle. In addition, glucose controlled flagella synthesis for the NPEC strain, but not for the hypermotile NPEC variant or the UPEC strain. The requirements for surface motility are likely to reflect major metabolic differences between strains for the pathways and regulation of energy metabolism.IMPORTANCEUrinary tract infections (UTIs) are one of the most common bacterial infections and are an increasing burden on the healthcare system because of recurrence and antibiotic resistance (1, 2). The most common uropathogen is E. coli (3, 4), which is responsible for about 80-90% of community acquired UTIs and 40-50% of nosocomial acquired UTIs (2). Virulence requires both pili and flagella, and either appendage can contribute to surface motility, although surface motility of uropathogenic E. coli has not been examined. We found different appendage, nutrient and pathway requirements for surface motility of a nonpathogenic E. coli lab strain and a uropathogenic E. coli We propose that these differences are the result of differences in the pathways and regulation of energy metabolism.