Abstract

The FeoABC ferrous transporter is a wide-spread bacterial system. While the feoABC locus is regulated by a number of factors in the bacteria studied, we have previously found that regulation of feoABC in Yersinia pestis appears to be unique. None of the non-iron responsive transcriptional regulators that control expression of feoABC in other bacteria do so in Y. pestis. Another unique factor is the iron and Fur regulation of the Y. pestis feoABC locus occurs during microaerobic but not aerobic growth. Here we show that this unique iron-regulation is not due to a unique aspect of the Y. pestis Fur protein but to DNA sequences that regulate transcription. We have used truncations, alterations, and deletions of the feoA::lacZ reporter to assess the mechanism behind the failure of iron to repress transcription under aerobic conditions. These studies plus EMSAs and DNA sequence analysis have led to our proposal that the feoABC locus has two promoters: an upstream P1 promoter whose expression is relatively iron-independent but repressed under microaerobic conditions and the known downstream Fur-regulated P2 promoter. In addition, we have identified two regions that bind Y. pestis protein(s), although we have not identified these protein(s) or their function. Finally we used iron uptake assays to demonstrate that both FeoABC and YfeABCD transport ferrous iron in an energy-dependent manner and also use ferric iron as a substrate for uptake.

Highlights

  • Iron is an essential nutrient and plays an important role in the pathogenesis of many organisms

  • KIM6 (∆pgm Feo+) strains carrying the various reporter plasmids were grown in chelex 100-treated PMH2 (cPMH2) in the presence and absence of iron under aerobic growth conditions at 37◦C

  • The FeoABC system is widespread in bacteria and transcriptional regulation by Fnr, ArcA, RstAB and Fur has been demonstrated, in E. coli, Salmonella, and Shigella (Kammler et al, 1993; Boulette and Payne, 2007; Jeon et al, 2008)

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Summary

Introduction

Iron is an essential nutrient and plays an important role in the pathogenesis of many organisms. The yersiniabactin (Ybt) siderophore system is one of the most important for obtaining ferric iron (Kirillina et al, 2006; Perry and Fetherston, 2011) and is critical for virulence during pneumonic and the early stages of bubonic plague (Bearden et al, 1997; Fetherston et al, 1999, 2010). The functions of 4 loci within the pgm locus have been determined: (1) ybt which encodes genes for the biosynthesis, regulation, and transport of the Ybt siderophore, (2) fet-flp, which encodes a ferrous iron transport system, (3) hms, which encodes components for biofilm development and (4) rip, which is involved in intracellular survival in phagocytic cells (Jarrett et al, 2004; Kirillina et al, 2004; Pujol et al, 2009; Forman et al, 2010)

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