1887

Abstract

The locus of enterocyte effacement (LEE) is a 35 kb pathogenicity island involved in attaching and effacing (A/E) enteric infection. The LEE is organized into five large transcriptional operons () and a few bi- and monocistronic instances. The operon co-transcribes three genes, , although can be transcribed in a separate mRNA from its own proximal promoter. The role of this separate promoter is not understood. In this study we characterized promoter activity for the type III secretion chaperone gene . The promoter, , has features consistent with σ promoters that contain an extended −10 element. This was experimentally confirmed by mutations that altered activity. In stark contrast to transcriptional operons, did not require the master regulator Ler for transcriptional activity. Moreover, activity was not dependent on the presence of GrlA or GrlR, two regulators associated with LEE gene expression. A fusion was used in real-time assays and demonstrated initial activity that occurred before promoter activity, which ensued after 120 min. was shown to be active during infection of HT-29 colonic epithelial cells. Inactivation of reduced CesT protein levels at early growth time points. These data indicate a Ler-, GrlA- and GrlR-independent activity for . We suggest that A/E pathogenic have evolved a mechanism to ready the cell for CesT protein expression in support of infection prior to Ler- and GrlA-related activities.

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2012-09-01
2020-12-05
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