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Chapter 3 : New Insights into Pheromone Control and Response in pCF10

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Abstract:

One of the best-studied forms of microbial behavior controlled by intercellular signaling is pheromone-inducible conjugation in . This chapter emphasizes significant results of research on the pheromone systems during the past several years, particularly the pCF10 system. The chapter focuses on new insights into the control of pheromone activity in donor and recipient cells, on the molecular mechanism of the pheromone response, and on the evolution of pCF10 plasmids. The majority of research on pCF10 plasmids has focused on pAD1 and pCF10. The genes encoded on the approximately 70-kb pCF10 plasmid are involved in pheromone-inducible conjugation. Specific outcomes of pheromone induction are discussed in the chapter. The authors describe the synthesis of cCF10 to exemplify a process that occurs in a similar fashion for several pheromones. The genetic screens and biochemical evidence demonstrating two functions for PrgX resulted in the development of a model of how PrgX negatively regulates from the prgQ promoter as well as positively regulates its own expression by a single mechanism. The X-ray crystal structure of PrgX is mainly helical with 17 alpha-helices. The authors propose that the pCF10 system described in the chapter has been appropriated by the plasmid for the purpose of increasing the sensitivity and adaptability to multiple situations.

Citation: Haemig H, Dunny G. 2008. New Insights into Pheromone Control and Response in pCF10, p 31-49. In Winans S, Bassler B (ed), Chemical Communication among Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815578.ch3

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Image of FIGURE 1
FIGURE 1

Model for pheromone-inducible conjugative transfer of pCF10. Recipient cell produces enough cCF10 pheromone to overcome iCF10 inhibitor produced by the donor cell. Pheromone cCF10 enters the donor cell through PrgZ and interacts with PrgX, leading to expression of the conjugation machinery including AS for plasmid transfer.

Citation: Haemig H, Dunny G. 2008. New Insights into Pheromone Control and Response in pCF10, p 31-49. In Winans S, Bassler B (ed), Chemical Communication among Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815578.ch3
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Image of FIGURE 2
FIGURE 2

A Proteolytic processing of cCF10 and iCF10. cCF10 is processed from CcfA in several steps. SPII, signal peptidase II; Eep, membrane protease; CP, carboxy peptidase. Arrows indicate cleavage sites. iCF10 is processed from a peptide produced from the 5′ end of the operon that is similar to the signal sequence of CcfA. B. Model for PrgY. The CcfA lipoprotein signal sequence is released from full-length polypeptide by SPII and further processed by Eep to generate cCF10. The primary functional activity of PrgY is to bind and sequester, degrade, or modify newly produced cCF10 as it is exported from the membrane following processing by Eep. In this model it is depicted that an interaction with Eep is mediated by the C-terminal intramembrane segments of PrgY. This would position PrgY optimally to intercept any newly released cCF10 from Eep.“N” depicts the amino terminus of the full-length CcfA polypeptide. The large extracellular domain of PrgY corresponds to the amino-terminal portion of the protein.

Citation: Haemig H, Dunny G. 2008. New Insights into Pheromone Control and Response in pCF10, p 31-49. In Winans S, Bassler B (ed), Chemical Communication among Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815578.ch3
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Image of FIGURE 3
FIGURE 3

Detailed map of the region. P and P are indicated by flags. Transcription from P results in two RNA products dependent on the induction state of the cell. Qs RNA encodes iCF10 and Q encodes a longer RNA important for readthrough to . RNA transcribed from P is processed to release Qa RNA, one negative regulator, and mRNA, which is translated to produce PrgX, the other negative regulatory molecule. Two binding sites for PrgX exist on the DNA, indicated by triple lines. The lollipop structures represent stem-loop structures formed in the message, with those downstream of P named IRS1 and IRS2.

Citation: Haemig H, Dunny G. 2008. New Insights into Pheromone Control and Response in pCF10, p 31-49. In Winans S, Bassler B (ed), Chemical Communication among Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815578.ch3
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Image of FIGURE 4
FIGURE 4

Structural consequences of inhibitor and pheromone binding on PrgX and the DNA looping model. (Top) Two PrgX molecules interlock via domain swapping of the N-terminal DNA-binding domain to form an X-shaped dimer. Dimers of PrgX complexed with iCF10 (black crosses) form a tetramer through interactions between the C-termini arms (black curls). PrgX tetramer increases affinity of each dimer for the DNA and is further stabilized by DNA looping. (Bottom) cCF10 occupation of the binding site causes the C-terminal arm (black line) to change structure and rotate to weaken interaction between dimers. The DNA unloops and the affinity of PrgX for the DNA decreases, allowing increased polymerase access to the promoter.

Citation: Haemig H, Dunny G. 2008. New Insights into Pheromone Control and Response in pCF10, p 31-49. In Winans S, Bassler B (ed), Chemical Communication among Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815578.ch3
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Image of FIGURE 5
FIGURE 5

Flow diagram indicating the state of RNAs in the uninduced and induced cell. In the uninduced state, transcripts from the P promoter are processed into mRNA and Qa RNA. Qa is a 102-nt RNA processed from the 5′ end and is complementary to the 3′ end of Qs RNA, an RNA constitutively produced at the promoter. It is predicted that interaction between Qa and Qs causes a folding such that Qs elongation terminates at IRS1, preventing readthrough to Q. Upon induction, the cellular levels of Qs greatly increase beyond those of Qa, resulting in unpaired Qs RNA. The unpaired Qs RNA does not terminate at IRS1 and is extended into Q and beyond, activating later steps in the induction process.

Citation: Haemig H, Dunny G. 2008. New Insights into Pheromone Control and Response in pCF10, p 31-49. In Winans S, Bassler B (ed), Chemical Communication among Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815578.ch3
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Image of FIGURE 6
FIGURE 6

Map of pCF10. Each predicted Orf of pCF10 derived from its complete sequence is indicated by a filled arrow ( ). Not shown is an approximately 16-kb segment consisting of the tetracycline-resistance element Tn which is not involved in pheromone-inducible conjugation. The evidence for independent evolution of the three modules indicated on the map is described in the text and in reference .

Citation: Haemig H, Dunny G. 2008. New Insights into Pheromone Control and Response in pCF10, p 31-49. In Winans S, Bassler B (ed), Chemical Communication among Bacteria. ASM Press, Washington, DC. doi: 10.1128/9781555815578.ch3
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References

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1. An, F. Y.,, M. C. Sulavik, and, D. B. Clewell. 1999. Identification and characterization of a determinant (eep) on the Enterococcus faecalis chromosome that is involved in production of the peptide sex pheromone cAD1. J. Bacteriol. 181:59155921.
2. Bae, T.,, S. Clerc-Bardin, and, G. M. Dunny. 2000. Analysis of expression of prgX, a key negative regulator of the transfer of the Enterococcus faecalis pheromone-inducible plasmid pCF10. J. Mol. Biol. 297:861875.
3. Bae, T.,, and G. M. Dunny. 2001. Dominant-negative mutants of prgX: evidence for a role for PrgX dimerization in negative regulation of pheromone-inducible conjugation. Mol. Microbiol. 39:13071320.
4. Bae, T.,, B. Kozlowicz, and, G. M. Dunny. 2002. Two targets in pCF10 DNA for PrgX binding: their role in production of Qa and prgX mRNA and in regulation of pheromone-inducible conjugation. J. Mol. Biol. 315:9951007.
5. Bae, T.,, B. K. Kozlowicz, and, G. M. Dunny. 2004. Characterization of cis-acting prgQ mutants: evidence for two distinct repression mechanisms by Qa RNA and PrgX protein in pheromoneinducible enterococcal plasmid pCF10. Mol. Microbiol. 51:271281.
6. Bensing, B. A.,, and G. M. Dunny. 1997. Pheromone-inducible expression of an aggregation protein in Enterococcus faecalis requires interaction of a plasmid-encoded RNA with components of the ribosome. Mol. Microbiol. 24:295308.
7. Brown, M. S.,, J. Ye,, R. B. Rawson, and, J. L. Goldstein. 2000. Regulated intramembrane proteolysis: a control mechanism conserved from bacteria to humans. Cell 100:391398.
8. Buttaro, B. A.,, M. H. Antiporta, and, G. M. Dunny. 2000. Cell-associated pheromone peptide (cCF10) production and pheromone inhibition in Enterococcus faecalis. J. Bacteriol. 182:49264933.
9. Chandler, J. R.,, and G. M. Dunny. 2004. Enterococcal peptide sex pheromones: synthesis and control of biological activity. Peptides 25:13771388.
10. Chandler, J. R.,, and G. M. Dunny. 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., in press.
11. Chandler, J. R.,, A. R. Flynn,, E. M. Bryan, and, G. M. Dunny. 2005. Specific control of endogenous cCF10 pheromone by a conserved domain of the pCF10-encoded regulatory protein PrgY in Enterococcus faecalis. J. Bacteriol. 187:48304843.
12. Chandler, J. R.,, H. Hirt, and, G. M. Dunny. 2005. A paracrine peptide sex pheromone also acts as an autocrine signal to induce plasmid transfer and virulence factor expression in vivo. Proc. Natl. Acad. Sci. USA 102:1561715622.
13. Chen, Y.,, J. H. Staddon, and, G. M. Dunny. 2007. Specificity determinants of conjugative DNA processing in the Enterococcus faecalis plasmid pCF10 and the Lactococcus lactis plasmid pRS01. Mol. Microbiol. 63:15491565.
14. Chow, J. W.,, L. A. Thal,, M. B. Perri,, J. A. Vazquez,, S. M. Donabedian,, D. B. Clewell, and, M. J. Zervos. 1993. Plasmid-associated hemolysin and aggregation substance production contribute to virulence in experimental enterococcal endocarditis. Antimicrob. Agents Chemother. 37:24742477.
15. Chung, J. W.,, and G. M. Dunny. 1995. Transcriptional analysis of a region of the Enterococcus faecalis plasmid pCF10 involved in positive regulation of conjugative transfer functions. J. Bacteriol. 177:21182124.
16. Clewell, D. B. 1999. Sex pheromone systems in enterococci, p. 47–65. In G. M. Dunny, and S. C. Winans (ed.), Cell-Cell Signaling in Bacteria. ASM Press, Washington, DC.
17. Clewell, D. B.,, and G. M. Dunny. 2002. Conjugation and genetic exchange in enterococci, p. 265–300. In D. B. Clewell, and M. S. Gilmore (ed.), The Enterococci: Pathogenesis, Molecular Biology, and Antibiotic Resistance. ASM Press, Washington, DC.
18. De Boever, E. H.,, D. B. Clewell, and, C. M. Fraser. 2000. Enterococcus faecalis conjugative plasmid pAM373: complete nucleotide sequence and genetic analyses of sex pheromone response. Mol. Microbiol. 37:13271341.
19. Dunny, G. M.,, M. H. Antiporta, and, H. Hirt. 2001. Peptide pheromone-induced transfer of plasmid pCF10 in Enterococcus faecalis: probing the genetic and molecular basis for specificity of the pheromone response. Peptides 22:15291539.
20. Dunny, G. M.,, B. L. Brown, and, D. B. Clewell. 1978. Induced cell aggregation and mating in Streptococcus faecalis: evidence for a bacterial sex pheromone. Proc. Natl. Acad. Sci. USA 75:34793483.
21. Dunny, G. M.,, and B. A. Leonard. 1997. Cell-cell communication in gram-positive bacteria. Annu. Rev. Microbiol. 51:527564.
22. Ehrenfeld, E. E.,, R. E. Kessler, and, D. B. Clewell. 1986. Identification of pheromone-induced surface proteins in Streptococcus faecalis and evidence of a role for lipoteichoic acid in formation of mating aggregates. J. Bacteriol. 168:612.
23. Fixen, K. R.,, J. R. Chandler,, T. Le,, B. K. Kozlowicz,, D. A. Manias, and, G. M. Dunny. 2006. Analysis of the amino acid sequence specificity determinants of the enterococcal cCF10 sex pheromone in the interactions with the pheromone-sensing machinery. J. Bacteriol. 189:13991406.
24. Francia, M. V.,, W. Haas,, R. Wirth,, E. Samberger,, A. Muscholl-Silberhorn,, M. S. Gilmore,, Y. Ike,, K. E. Weaver,, F. Y. An, and, D. B. Clewell. 2001. Completion of the nucleotide sequence of the Enterococcus faecalis conjugative virulence plasmid pAD1 and identification of a second transfer origin. Plasmid 46:117127.
25. Fujimoto, S.,, M. Bastos,, K. Tanimoto,, F. An,, K. Wu, and, D. B. Clewell. 1997. The pAD1 sex pheromone response in Enterococcus faecalis. Adv. Exp. Med. Biol. 418:10371040.
26. Fujimoto, S.,, and D. B. Clewell. 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 95:64306435.
27. Hedberg, P. J.,, B. A. Leonard,, R. E. Ruhfel, and, G. M. Dunny. 1996. Identification and characterization of the genes of Enterococcus faecalis plasmid pCF10 involved in replication and in negative control of pheromone-inducible conjugation. Plasmid 35:4657.
28. Hirt, H.,, D. A. Manias,, E. M. Bryan,, J. R. Klein,, J. K. Marklund,, J. H. Staddon,, M. L. Paustian,, V. Kapur, and, G. M. Dunny. 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. 187:10441054.
29. Hirt, H.,, P. M. Schlievert, and, G. M. Dunny. 2002. In vivo induction of virulence and antibiotic resistance transfer in Enterococcus faecalis mediated by the sex pheromone-sensing system of pCF10. Infect. Immun. 70:716723.
30. Horii, T.,, H. Nagasawa, and, J. Nakayama. 2002. Functional analysis of TraA, the sex pheromone receptor encoded by pPD1, in a promoter region essential for the mating response in Enterococcus faecalis. J. Bacteriol. 184:63436350.
31. Hu, J. C. 1995. Repressor fusions as a tool to study protein-protein interactions. Structure 3:431433.
32. Isenmann, R.,, M. Schwarz,, E. Rozdzinski,, R. Marre, and, H. G. Beger. 2000. Aggregation substance promotes colonic mucosal invasion of Enterococcus faecalis in an ex vivo model. J. Surg. Res. 89:132138.
33. Kao, S. M.,, S. B. Olmsted,, A. S. Viksnins,, J. C. Gallo, and, G. M. Dunny. 1991. Molecular and genetic analysis of a region of plasmid pCF10 containing positive control genes and structural genes encoding surface proteins involved in pheromoneinducible conjugation in Enterococcus faecalis. J. Bacteriol. 173:76507664.
34. Kozlowicz, B. K.,, T. Bae, and, G. M. Dunny. 2004. Enterococcus faecalis pheromone-responsive protein PrgX: genetic separation of positive autoregulatory functions from those involved in negative regulation of conjugative plasmid transfer. Mol. Microbiol. 54:520532.
35. Kozlowicz, B. K.,, M. Dworkin, and, G. M. Dunny. 2006. Pheromone-inducible conjugation in Enterococcus faecalis: a model for the evolution of biological complexity? Int. J. Med. Microbiol. 296:141147.
36. Kozlowicz, B. K.,, K. Shi,, Z. Y. Gu,, D. H. Ohlendorf,, C. A. Earhart, and, G. M. Dunny. 2006. Molecular basis for control of conjugation by bacterial pheromone and inhibitor peptides. Mol. Microbiol. 62:958969.
37. Kreft, B.,, R. Marre,, U. Schramm, and, R. Wirth. 1992. Aggregation substance of Enterococcus faecalis mediates adhesion to cultured renal tubular cells. Infect. Immun. 60:2530.
38. Leonard, B. A.,, A. Podbielski,, P. J. Hedberg, and, G. M. Dunny. 1996. Enterococcus faecalis pheromone binding protein, PrgZ, recruits a chromosomal oligopeptide permease system to import sex pheromone cCF10 for induction of conjugation. Proc. Natl. Acad. Sci. USA 93:260264.
39. McCormick, J. K.,, H. Hirt,, C. M. Waters,, T. J. Tripp,, G. M. Dunny, and, P. M. Schlievert. 2001. Antibodies to a surface-exposed, N-terminal domain of aggregation substance are not protective in the rabbit model of Enterococcus faecalis infective endocarditis. Infect. Immun. 69:33053314.
40. McCormick, J. K.,, T. J. Tripp,, G. M. Dunny, and, P. M. Schlievert. 2002. Formation of vegetations during infective endocarditis excludes binding of bacterial-specific host antibodies to Enterococcus faecalis. J. Infect. Dis. 185:994997.
41. Mori, M.,, H. Tanaka,, Y. Sakagami,, A. Isogai,, M. Fujino,, C. Kitada,, D. B. Clewell, and, A. Suzuki. 1987. Isolation and structure of the sex pheromone inhibitor, iPD1, excreted by Streptococcus faecalis donor strains harboring plasmid pPD1. J. Bacteriol. 169:17471749.
42. Muscholl-Silberhorn, A. B. 2000. Pheromone-regulated expression of sex pheromone plasmid pAD1-encoded aggregation substance depends on at least six upstream genes and a cis-acting, orientation-dependent factor. J. Bacteriol. 182:38163825.
43. Nakayama, J.,, R. E. Ruhfel,, G. M. Dunny,, A. Isogai, and, A. Suzuki. 1994. The prgQ gene of the Enterococcus faecalis tetracycline resistance plasmid pCF10 encodes a peptide inhibitor, iCF10. J. Bacteriol. 176:74057408.
44. Nakayama, J.,, Y. Takanami,, T. Horii,, S. Sakuda, and, A. Suzuki. 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. 180:449456.
45. Olmsted, S. B.,, G. M. Dunny,, S. L. Erlandsen, and, C. L. Wells. 1994. A plasmid-encoded surface protein on Enterococcus faecalis augments its internalization by cultured intestinal epithelial cells. J. Infect. Dis. 170:15491556.
46. Olmsted, S. B.,, S. M. Kao,, L. J. van Putte,, J. C. Gallo, and, G. M. Dunny. 1991. Role of the pheromone-inducible surface protein Asc10 in mating aggregate formation and conjugal transfer of the Enterococcus faecalis plasmid pCF10. J. Bacteriol. 173:76657672.
47. Ozawa, Y.,, E. H. De Boever, and, D. B. Clewell. 2005. Enterococcus faecalis sex pheromone plasmid pAM373: analyses of TraA and evidence for its interaction with RpoB. Plasmid 54:5769.
48. Rakita, R. M.,, N. N. Vanek,, K. Jacques-Palaz,, M. Mee,, M. M. Mariscalco,, G. M. Dunny,, M. Snuggs,, W. B. Van Winkle, and, S. I. Simon. 1999. Enterococcus faecalis bearing aggregation substance is resistant to killing by human neutrophils despite phagocytosis and neutrophil activation. Infect. Immun. 67:60676075.
49. Ruhfel, R. E.,, D. A. Manias, and, G. M. Dunny. 1993. Cloning and characterization of a region of the Enterococcus faecalis conjugative plasmid, pCF10, encoding a sex pheromone-binding function. J. Bacteriol. 175:52535259.
50. Sartingen, S.,, E. Rozdzinski,, A. Muscholl-Silberhorn, and, R. Marre. 2000. Aggregation substance increases adherence and internalization, but not translocation, of Enterococcus faecalis through different intestinal epithelial cells in vitro. Infect. Immun. 68:60446047.
51. Schlievert, P. M.,, P. J. Gahr,, A. P. Assimacopoulos,, M. M. Dinges,, J. A. Stoehr,, J. W. Harmala,, H. Hirt, and, G. M. Dunny. 1998. Aggregation and binding substances enhance pathogenicity in rabbit models of Enterococcus faecalis endocarditis. Infect. Immun. 66:218223.
52. Shi, K.,, C. K. Brown,, Z. Y. Gu,, B. K. Kozlowicz,, G. M. Dunny,, D. H. Ohlendorf, and, C. A. Earhart. 2005. Structure of peptide sex pheromone receptor PrgX and PrgX/pheromone complexes and regulation of conjugation in Enterococcus faecalis. Proc. Natl. Acad. Sci. USA 102:1859618601.
53. Staddon, J. H.,, E. M. Bryan,, D. A. Manias, and, G. M. Dunny. 2004. Conserved target for group II intron insertion in relaxase genes of conjugative elements of gram-positive bacteria. J. Bacteriol. 186:23932401.
54. Sussmuth, S. D.,, A. Muscholl-Silberhorn,, R. Wirth,, M. Susa,, R. Marre, and, E. Rozdzinski. 2000. Aggregation substance promotes adherence, phagocytosis, and intracellular survival of Enterococcus faecalis within human macrophages and suppresses respiratory burst. Infect. Immun. 68:49004906.
55. Tanimoto, K.,, and D. B. Clewell. 1993. Regulation of the pAD1-encoded sex pheromone response in Enterococcus faecalis: expression of the positive regulator TraE1. J. Bacteriol. 175:10081018.
56. Tanimoto, K.,, H. Tomita, and, Y. Ike. 1996. The traA gene of the Enterococcus faecalis conjugative plasmid pPD1 encodes a negative regulator for the pheromone response. Plasmid 36:5561.
57. Trotter, K. M.,, and G. M. Dunny. 1990. Mutants of Enterococcus faecalis deficient as recipients in mating with donors carrying pheromoneinducible plasmids. Plasmid 24:5767.
58. Waters, C. M.,, and G. M. Dunny. 2001. Analysis of functional domains of the Enterococcus faecalis pheromone-induced surface protein aggregation substance. J. Bacteriol. 183:56595667.
59. Waters, C. M.,, H. Hirt,, J. K. McCormick,, P. M. Schlievert,, C. L. Wells, and, G. M. Dunny. 2004. An amino-terminal domain of Enterococcus faecalis aggregation substance is required for aggregation, bacterial internalization by epithelial cells and binding to lipoteichoic acid. Mol. Microbiol. 52:11591171.
60. Waters, C. M.,, C. L. Wells, and, G. M. Dunny. 2003. The aggregation domain of aggregation substance, not the RGD motifs, is critical for efficient internalization by HT-29 enterocytes. Infect. Immun. 71:56825689.
61. Winans, S. C.,, D. L. Burns, and, P. J. Christie. 1996. Adaptation of a conjugal transfer system for the export of pathogenic macromolecules. Trends Microbiol. 4:6468.

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