This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Pomerantsev, A. P. Articles by Leppla, S. H. Search for Related Content PubMed PubMed Citation Articles by Pomerantsev, A. P. Articles by Leppla, S. H.

 Previous Article  |  Next Article 

Infection and Immunity, October 2004, p. 5814-5823, Vol. 72, No. 10
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.10.5814-5823.2004

A Spontaneous Translational Fusion of Bacillus cereus PlcR and PapR Activates Transcription of PlcR-Dependent Genes in Bacillus anthracis via Binding with a Specific Palindromic Sequence

Andrei P. Pomerantsev,¹ Olga M. Pomerantseva,² and Stephen H. Leppla^1*

Microbial Pathogenesis Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda,¹ Biological Defense Research Directorate, Naval Medical Research Center, Rockville, Maryland²

Received 20 April 2004/ Returned for modification 21 May 2004/ Accepted 7 July 2004

Transformation of Bacillus anthracis with plasmid pUTE29-plcR-papR carrying the native Bacillus cereus plcR-papR gene cluster did not activate expression of B. anthracis hemolysin genes, even though these are expected to be responsive to activation by the global regulator PlcR. To further characterize the action of PlcR, we examined approximately 3,000 B. anthracis transformants containing pUTE29-plcR-papR and found a single hemolytic colony. The hemolytic strain contained a plasmid having a spontaneous plcR-papR intergenic region deletion. Transformation of the resulting plasmid pFP12, encoding a fused PlcR-PapR protein, into the nonhemolytic B. anthracis parental strain produced strong activation of B. anthracis hemolysins, including phosphatidylcholine-specific phospholipase C and sphingomyelinase. The fused PlcR-PapR protein present in a lysate of B. anthracis containing pFP12 bound strongly and specifically to the double-stranded palindrome 5'-TATGCATTATTTCATA-3' that matches the consensus PlcR-binding site. In contrast, native PlcR protein in a lysate from a B. anthracis strain expressing large amounts of this protein did not demonstrate binding with the palindrome. The results suggest that the activation of PlcR by binding of a PapR pentapeptide as normally occurs in Bacillus thuringiensis and B. cereus can be mimicked by tethering the peptide to PlcR in a translational fusion, thereby obviating the need for PapR secretion, extracellular processing, retrieval into the bacterium, and binding with PlcR.

---

* Corresponding author. Mailing address: Microbial Pathogenesis Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-4350. Phone: (301) 594-2865. Fax: (301) 480-0326. E-mail: sleppla{at}niaid.nih.gov.

Editor: J. T. Barbieri

---

Infection and Immunity, October 2004, p. 5814-5823, Vol. 72, No. 10
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.10.5814-5823.2004

This article has been cited by other articles:

• Bouillaut, L., Perchat, S., Arold, S., Zorrilla, S., Slamti, L., Henry, C., Gohar, M., Declerck, N., Lereclus, D. (2008). Molecular basis for group-specific activation of the virulence regulator PlcR by PapR heptapeptides. Nucleic Acids Res 36: 3791-3801 [Abstract] [Full Text]  
• Banerjee, A., Biswas, I. (2008). Markerless Multiple-Gene-Deletion System for Streptococcus mutans. Appl. Environ. Microbiol. 74: 2037-2042 [Abstract] [Full Text]  
• Declerck, N., Bouillaut, L., Chaix, D., Rugani, N., Slamti, L., Hoh, F., Lereclus, D., Arold, S. T. (2007). Structure of PlcR: Insights into virulence regulation and evolution of quorum sensing in Gram-positive bacteria. Proc. Natl. Acad. Sci. USA 104: 18490-18495 [Abstract] [Full Text]  
• Miyakoshi, M., Shintani, M., Terabayashi, T., Kai, S., Yamane, H., Nojiri, H. (2007). Transcriptome Analysis of Pseudomonas putida KT2440 Harboring the Completely Sequenced IncP-7 Plasmid pCAR1. J. Bacteriol. 189: 6849-6860 [Abstract] [Full Text]  
• Ross, C. L., Koehler, T. M. (2006). plcR papR-Independent Expression of Anthrolysin O by Bacillus anthracis. J. Bacteriol. 188: 7823-7829 [Abstract] [Full Text]  
• Wei, Z., Schnupf, P., Poussin, M. A., Zenewicz, L. A., Shen, H., Goldfine, H. (2005). Characterization of Listeria monocytogenes Expressing Anthrolysin O and Phosphatidylinositol-Specific Phospholipase C from Bacillus anthracis. Infect. Immun. 73: 6639-6646 [Abstract] [Full Text]