This Article 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 Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ruhfel, R E Articles by Dunny, G M Search for Related Content PubMed PubMed Citation Articles by Ruhfel, R E Articles by Dunny, G M

research-article

Cloning and characterization of a region of the Enterococcus faecalis conjugative plasmid, pCF10, encoding a sex pheromone-binding function.

Institute for Advanced Studies in Biological Process Technology, University of Minnesota, St. Paul 55108.

ABSTRACT

In order to investigate the mechanism by which peptide sex pheromones induce expression of the conjugation functions of certain Enterococcus faecalis plasmids, a biological assay was developed to measure the ability of cells carrying the conjugative plasmid pCF10 to bind the sex pheromone cCF10. The data indicated that pCF10 endows its host E. faecalis cell with the ability to specifically remove (apparently by irreversible binding) cCF10 activity from culture medium. The pCF10 DNA encoding this ability was localized to a 3.4-kb segment within a region involved in negative control of expression of conjugal transfer functions. This segment also encoded ability to bind the pheromone inhibitor peptide iCF10. DNA sequencing revealed three open reading frames, which have been denoted prgW (pheromone responsive gene W), prgZ, and prgY. The deduced product of prgW resembled regulatory proteins from other bacteria and eucaryotes, with a very high degree of identity within a putative DNA-binding domain. The prgY gene actually extended into an adjacent region of pCF10 and could encode a protein with significant similarity to a protein called TraB, believed to be involved in shutdown of pheromone cAD1 production by cells carrying the pheromone-inducible hemolysin plasmid pAD1, according to F.Y. An and D.B. Clewell (Abstr. Gen. Meet. Am. Soc. Microbiol. 1992, H70, 1992). The prgZ gene product showed significant relatedness to binding proteins encoded by oligopeptide permease (opp) operons in gram-positive and gram-negative bacteria and is highly similar to a pAD1-encoded protein, TraC, which is believed to mediate sex pheromone cAD1 binding (K. Tanimoto, F. Y. An, and D. B. Clewell, submitted for publication). A Tn5 insertion into prgZ abolished cCF10 binding ability.

This article has been cited by other articles:

• Chandler, J. R., Dunny, G. M. (2008). Characterization of the Sequence Specificity Determinants Required for Processing and Control of Sex Pheromone by the Intramembrane Protease Eep and the Plasmid-Encoded Protein PrgY. J. Bacteriol. 190: 1172-1183 [Abstract] [Full Text]  
• Dufour, M., Manson, J. M., Bremer, P. J., Dufour, J.-P., Cook, G. M., Simmonds, R. S. (2007). Characterization of Monolaurin Resistance in Enterococcus faecalis. Appl. Environ. Microbiol. 73: 5507-5515 [Abstract] [Full Text]  
• Chandler, J. R., Flynn, A. R., Bryan, E. M., Dunny, G. M. (2005). Specific Control of Endogenous cCF10 Pheromone by a Conserved Domain of the pCF10-Encoded Regulatory Protein PrgY in Enterococcus faecalis. J. Bacteriol. 187: 4830-4843 [Abstract] [Full Text]  
• Hirt, H., Manias, D. A., Bryan, E. M., Klein, J. R., Marklund, J. K., Staddon, J. H., Paustian, M. L., Kapur, V., Dunny, G. M. (2005). Characterization of the Pheromone Response of the Enterococcus faecalis Conjugative Plasmid pCF10: Complete Sequence and Comparative Analysis of the Transcriptional and Phenotypic Responses of pCF10-Containing Cells to Pheromone Induction. J. Bacteriol. 187: 1044-1054 [Abstract] [Full Text]  
• Sanchez-Hidalgo, M., Maqueda, M., Galvez, A., Abriouel, H., Valdivia, E., Martinez-Bueno, M. (2003). The Genes Coding for Enterocin EJ97 Production by Enterococcus faecalis EJ97 Are Located on a Conjugative Plasmid. Appl. Environ. Microbiol. 69: 1633-1641 [Abstract] [Full Text]  
• Hirt, H., Schlievert, P. M., Dunny, G. M. (2002). In Vivo Induction of Virulence and Antibiotic Resistance Transfer in Enterococcus faecalis Mediated by the Sex Pheromone-Sensing System of pCF10. Infect. Immun. 70: 716-723 [Abstract] [Full Text]  
• Buttaro, B. A. L., Antiporta, M. H., Dunny, G. M. (2000). Cell-Associated Pheromone Peptide (cCF10) Production and Pheromone Inhibition in Enterococcus faecalis. J. Bacteriol. 182: 4926-4933 [Abstract] [Full Text]  
• Firth, N., Apisiridej, S., Berg, T., O'Rourke, B. A., Curnock, S., Dyke, K. G. H., Skurray, R. A. (2000). Replication of Staphylococcal Multiresistance Plasmids. J. Bacteriol. 182: 2170-2178 [Abstract] [Full Text]  
• Kornacki, J. A., Oliver, D. B. (1998). Lyme Disease-Causing Borrelia Species Encode Multiple Lipoproteins Homologous to Peptide-Binding Proteins of ABC-Type Transporters. Infect. Immun. 66: 4115-4122 [Abstract] [Full Text]  
• Berg, T., Firth, N., Apisiridej, S., Hettiaratchi, A., Leelaporn, A., Skurray, R. A. (1998). Complete Nucleotide Sequence of pSK41: Evolution of Staphylococcal Conjugative Multiresistance Plasmids. J. Bacteriol. 180: 4350-4359 [Abstract] [Full Text]  
• Fujimoto, S., Clewell, D. B. (1998). Regulation of the pAD1 sex pheromone response of Enterococcus faecalis by direct interaction between the cAD1 peptide mating signal and the negatively regulating, DNA-binding TraA protein. Proc. Natl. Acad. Sci. USA 95: 6430-6435 [Abstract] [Full Text]  
• Nakayama, J., Takanami, Y., Horii, T., Sakuda, S., Suzuki, A. (1998). Molecular Mechanism of Peptide-Specific Pheromone Signaling in Enterococcus faecalis: Functions of Pheromone Receptor TraA and Pheromone-Binding Protein TraC Encoded by Plasmid pPD1. J. Bacteriol. 180: 449-456 [Abstract] [Full Text]  
• Solomon, J M, Magnuson, R, Srivastava, A, Grossman, A D (1995). Convergent sensing pathways mediate response to two extracellular competence factors in Bacillus subtilis.. Genes Dev. 9: 547-558 [Abstract]