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Journal of Bacteriology, February 2007, p. 1399-1406, Vol. 189, No. 4
0021-9193/07/$08.00+0     doi:10.1128/JB.01226-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Analysis of the Amino Acid Sequence Specificity Determinants of the Enterococcal cCF10 Sex Pheromone in Interactions with the Pheromone-Sensing Machinery{triangledown}

Kathryn R. Fixen,{dagger} Josephine R. Chandler,{ddagger} Thinh Le, Briana K. Kozlowicz,§ Dawn A. Manias, and Gary M. Dunny*

Department of Microbiology, University of Minnesota, MMC 196, 420 Delaware Street, S.E., Minneapolis, Minnesota 55455

Received 4 August 2006/ Accepted 2 November 2006

The level of expression of conjugation genes in Enterococcus faecalis strains carrying the pheromone-responsive transferable plasmid pCF10 is determined by the ratio in the culture medium of two types of signaling peptides, a pheromone (cCF10) and an inhibitor (iCF10). Recent data have demonstrated that both peptides target the cytoplasmic receptor protein PrgX. However, the relative importance of the interaction of these peptides with the pCF10 protein PrgZ (which enhances import of cCF10) versus PrgX is not fully understood, and there is relatively little information about specific amino acid sequence determinants affecting the functional interactions of cCF10 with these proteins in vivo. To address these issues, we used a pheromone-inducible reporter gene system where various combinations of PrgX and PrgZ could be expressed in an isogenic host background to examine the biological activities of cCF10, iCF10, and variants of cCF10 isolated in a genetic screen. The results suggest that most of the amino acid sequence determinants of cCF10 pheromone activity affect interactions between the peptide and PrgX, although some sequence variants that affected peptide/PrgZ interactions were also identified. The results provide functional data to complement ongoing structural studies of PrgX and increase our understanding of the functional interactions of cCF10 and iCF10 with the pheromone-sensing machinery of pCF10.


* Corresponding author. Mailing address: Department of Microbiology, University of Minnesota, MMC 196, 420 Delaware St., S.E., Minneapolis, MN 55455. Phone: (612) 625-9930. Fax: (612) 626-0623. E-mail: dunny001{at}umn.edu.

{triangledown} Published ahead of print on 10 November 2006.

{dagger} Present address: Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA.

{ddagger} Present address: Department of Microbiology, University of Washington, Seattle, WA.

§ Present address: Oregon Health and Science University, Department of Molecular Microbiology and Immunology, Portland, OR.


Journal of Bacteriology, February 2007, p. 1399-1406, Vol. 189, No. 4
0021-9193/07/$08.00+0     doi:10.1128/JB.01226-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.




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]