1887

Abstract

Uropathogenic (UPEC) causes more than 90 % of all human urinary tract infections through type 1 piliated UPEC cells binding to bladder epithelial cells. The FimB and FimE site-specific recombinases orient the element containing the structural gene promoter. Regulation of and depends on environmental pH and osmolality. The EnvZ/OmpR two-component system affects osmoregulation in . To ascertain if OmpR directly regulated the gene promoters, gel mobility shift and DNase I footprinting experiments were performed using OmpR or phosphorylated OmpR (OmpR-P) mixed with the promoter regions of UPEC strain NU149. Both OmpR-P and OmpR bound weakly to one promoter. Because there was weak binding to one promoter, strain NU149 was grown in different pH and osmolality environments, and total RNAs were extracted from each population and converted to cDNAs. Quantitative reverse-transcriptase PCR showed no differences in transcription among the different growth conditions. Conversely, Western blots showed a significant increase in OmpR protein in UPEC cells grown in a combined low pH/high osmolality environment versus a neutral pH/high osmolality environment. In a high osmolality environment, the mutant expressed more transcripts and Phase-ON positioning of the element as well as higher type 1 pili levels than wild-type cells. Together these results suggest that OmpR may be post-transcriptionally regulated in UPEC cells growing in a low pH/high osmolality environment, which regulates in UPEC.

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2013-02-01
2020-01-27
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