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Infection and Immunity, December 2001, p. 7772-7782, Vol. 69, No. 12
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.12.7772-7782.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Expression Analysis of the Yersiniabactin Receptor Gene fyuA and the Heme Receptor hemR of Yersinia enterocolitica In Vitro and In Vivo Using the Reporter Genes for Green Fluorescent Protein and Luciferase

Christoph A. Jacobi, Sebastian Gregor, Alexander Rakin, and Jürgen Heesemann*

Max von Pettenkofer Institute for Hygiene and Medical Microbiology, Ludwig Maximillian University Munich, Munich, Germany

Received 4 June 2001/Returned for modification 9 August 2001/Accepted 19 September 2001

The enteropathogenic Yersinia enterocolitica strains have several systems for scavenging iron from their environment. We have studied the expression of the fyuA gene, which encodes the outer membrane receptor for the siderophore yersiniabactin (Ybt), and the hemR gene, which encodes the receptor for heme, using the reporter genes gfp (encoding green fluorescent protein) and luc (encoding firefly luciferase). To study gene expression in vitro as well as in vivo, we have constructed several translational reporter gene fusions to monitor simultaneously expression of fyuA and hemR or expression of one gene by a gfp-luc tandem reporter. Results of the in vitro expression analysis (liquid media) indicated that fyuA and hemR are strongly derepressed under iron starvation conditions, resulting in strong fluorescence and/or luminescence at 27°C. In the in vivo BALB/C mouse infection model, tissue-specific expression of fyuA and hemR reporter fusions was observed. Surprisingly, fyuA and hemR reporter constructs were weakly expressed by yersiniae located in the liver and intestinal lumen, whereas strong expression was found for yersiniae in the peritoneal cavity and moderate expression was found in the spleen. Strikingly, yersiniae carrying fyuA or hemR reporter fusions exhibited threefold-stronger signals when grown in the peritoneal cavity of mice than those growing under iron derepression in vitro. This hyperexpression suggests that besides Fur derepression, additional activators may be involved in the enhanced expression of fyuA and hemR under peritoneal growth conditions. Differential expression of the fyuA and hemR reporter fusions could not be observed, suggesting similar regulation of fyuA and hemR in the mouse infection model.

* Corresponding author. Mailing address: Max von Pettenkofer Institut for Hygiene and Medical Microbiology, Ludwig Maximillian University München, Pettenkoferstrasse 9a, 80336 München, Germany. Phone: (49-89) 51605200. Fax: (49-89) 51605202. E-mail: heesemann{at}m3401.mpk.med.uni-muenchen.de.

Infection and Immunity, December 2001, p. 7772-7782, Vol. 69, No. 12
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.12.7772-7782.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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