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Journal of Bacteriology, November 2006, p. 7941-7956, Vol. 188, No. 22
0021-9193/06/$08.00+0     doi:10.1128/JB.00473-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Species-Specific Differences in the Activity of PrfA, the Key Regulator of Listerial Virulence Genes{triangledown}

Norman Mauder,1,{dagger} Regina Ecke,1,{dagger} Sonja Mertins,1 Daniela I. M. Loeffler,1 Gerald Seidel,2 Mareen Sprehe,2 Wolfgang Hillen,2 Werner Goebel,1* and Stefanie Müller-Altrock1

Lehrstuhl für Mikrobiologie, Biozentrum, Universität Würzburg, D-97074 Würzburg, Germany,1 Lehrstuhl für Mikrobiologie, Institut für Mikrobiologie, Biochemie und Genetik der Friedrich-Alexander Universität Erlangen-Nürnberg, D-91058, Erlangen, Germany2

Received 5 April 2006/ Accepted 5 September 2006

PrfA, the master regulator of LIPI-1, is indispensable for the pathogenesis of the human pathogen Listeria monocytogenes and the animal pathogen Listeria ivanovii. PrfA is also present in the apathogenic species Listeria seeligeri, and in this study, we elucidate the differences between PrfA proteins from the pathogenic and apathogenic species of the genus Listeria. PrfA proteins of L. monocytogenes (PrfALm and PrfA*Lm), L. ivanovii (PrfALi), and L. seeligeri (PrfALs) were purified, and their equilibrium constants for binding to the PrfA box of the hly promoter (PhlyLm) were determined by surface plasmon resonance. In addition, the capacities of these PrfA proteins to bind to the PrfA-dependent promoters Phly and PactA and to form ternary complexes together with RNA polymerase were analyzed in electrophoretic mobility shift assays, and their abilities to initiate transcription in vitro starting at these promoters were compared. The results show that PrfALi resembled the constitutively active mutant PrfA*Lm more than the wild-type PrfALm, whereas PrfALs showed a drastically reduced capacity to bind to the PrfA-dependent promoters Phly and PactA. In contrast, the efficiencies of PrfALm, PrfA*Lm, and PrfALi forming ternary complexes and initiating transcription at Phly and PactA were rather similar, while those of PrfALs were also much lower. The low binding and transcriptional activation capacities of PrfALs seem to be in part due to amino acid exchanges in its C-terminal domain (compared to PrfALm and PrfALi). In contrast to the significant differences in the biochemical properties of PrfALm, PrfALi, and PrfALs, the PrfA-dependent promoters of hly (PhlyLm, PhlyLi, and PhlyLs) and actA (PactALm, PactALi, and PactALs) of the three Listeria species did not significantly differ in their binding affinities to the various PrfA proteins and in their strengths to promote transcription in vitro. The allelic replacement of prfALm with prfALs in L. monocytogenes leads to low expression of PrfA-dependent genes and to reduced in vivo virulence of L. monocytogenes, suggesting that the altered properties of PrfALs protein are a major cause for the low virulence of L. seeligeri.

* Corresponding author. Mailing address: Lehrstuhl für Mikrobiologie, Biozentrum, Universität Würzburg, Am Hubland, D-97074 Würzburg, Germany. Phone: 49-931-8884401. Fax: 49-931-8884402. E-mail: goebel{at}biozentrum.uni-wuerzburg.de.

{triangledown} Published ahead of print on 15 September 2006.

{dagger} N.M. and R.E. contributed equally to this work.

Journal of Bacteriology, November 2006, p. 7941-7956, Vol. 188, No. 22
0021-9193/06/$08.00+0     doi:10.1128/JB.00473-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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  • Miner, M. D., Port, G. C., Freitag, N. E. (2008). Functional impact of mutational activation on the Listeria monocytogenes central virulence regulator PrfA. Microbiology 154: 3579-3589 [Abstract] [Full Text]  
  • Joseph, B., Mertins, S., Stoll, R., Schar, J., Umesha, K. R., Luo, Q., Muller-Altrock, S., Goebel, W. (2008). Glycerol Metabolism and PrfA Activity in Listeria monocytogenes. J. Bacteriol. 190: 5412-5430 [Abstract] [Full Text]  


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