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Research Article

The vfr gene product, required for Pseudomonas aeruginosa exotoxin A and protease production, belongs to the cyclic AMP receptor protein family.

S E West, A K Sample, L J Runyen-Janecky
S E West
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison 53706.
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A K Sample
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison 53706.
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L J Runyen-Janecky
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison 53706.
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DOI: 10.1128/jb.176.24.7532-7542.1994
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ABSTRACT

The synthesis of exotoxin A (ETA) by Pseudomonas aeruginosa is a complex, regulated event. Several ETA putative regulatory mutants of P. aeruginosa PA103 have previously been characterized (S. E. H. West, S. A. Kaye, A. N. Hamood, and B. H. Iglewski, Infect. Immun. 62:897-903, 1994). In addition to ETA production, these mutants, PA103-15, PA103-16, and PA103-19, were also deficient in the production of protease and in regA P1 promoter activity. RegA is a positive regulator of ETA transcription. We cloned a gene, designated vfr for virulence factor regulator, that restored ETA and protease production to parental levels in these mutants. In addition, transcription from the regA P1 promoter was restored. In Escherichia coli, when vfr was overexpressed from a phage T7 promoter, a protein with an apparent molecular mass of 28.5 kDa was produced. Analysis of the deduced amino acid sequence of vfr revealed that the expected protein is 67% identical and 91% similar over a 202-amino-acid overlap to the E. coli cyclic AMP receptor protein (CAP or Crp). The cloned vfr gene complemented the beta-galactosidase- and tryptophanase-deficient phenotypes of E. coli RZ1331, a crp deletion mutant. However, the E. coli crp gene under the control of the tac promoter did not complement the ETA-deficient or protease-deficient phenotype of PA103-15 or PA103-16. The ability of vfr to restore both ETA and protease production to these mutants suggests that vfr is a global regulator of virulence factor expression in P. aeruginosa.

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The vfr gene product, required for Pseudomonas aeruginosa exotoxin A and protease production, belongs to the cyclic AMP receptor protein family.
S E West, A K Sample, L J Runyen-Janecky
Journal of Bacteriology Dec 1994, 176 (24) 7532-7542; DOI: 10.1128/jb.176.24.7532-7542.1994

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The vfr gene product, required for Pseudomonas aeruginosa exotoxin A and protease production, belongs to the cyclic AMP receptor protein family.
S E West, A K Sample, L J Runyen-Janecky
Journal of Bacteriology Dec 1994, 176 (24) 7532-7542; DOI: 10.1128/jb.176.24.7532-7542.1994
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