1887

Abstract

Components of the bacterial phosphoenolpyruvate (PEP) : carbohydrate phosphortransferase system (PTS) have multiple regulatory roles in addition to PEP-dependent transport/phosphorylation of numerous carbohydrates. We have recently shown that, in an opportunistic human pathogen, , enzyme IIA (EIIA) interacts with a peptidase that has high sequence similarity to mammalian insulin-degrading enzymes, called insulin-degrading enzyme (vIDE). Although the vIDE–EIIA interaction is independent of the phosphorylation state of EIIA, vIDE shows no peptidase activity unless complexed with the unphosphorylated form of EIIA. A deletion mutant of , the gene encoding vIDE, shows remarkably lower degrees of survival and virulence than the wild-type strain in mice, implying that vIDE is a virulence factor. In this study, we investigated regulation of expression at the transcriptional level. Primer extension analysis identified two different transcriptional start sites of : P for the longer transcript and P for the shorter transcript. We performed ligand fishing experiments by using the promoter region of and found that the cAMP receptor protein (CRP) specifically binds to the promoter. DNase I footprinting experiments revealed that CRP binds to a region between the two promoters. transcription assays showed that CRP activates P transcription in the presence of cAMP whose concentration is regulated by EIIA. These results suggest that EIIA regulates the expression level of vIDE as well as its activity.

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2012-05-01
2019-10-15
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