Regulation of the pAD1-encoded sex pheromone response in Enterococcus faecalis: nucleotide sequence analysis of traA

Pontius, Linda T.; Clewell, Don B.

doi:10.1128/jb.174.6.1821-1827.1992

Cited by 39 publications

(39 citation statements)

References 42 publications

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“…The deduced protein encoded by traA exhibited significant homology with TraA (36.6%) of pAD1 (35) and PrgX (21%) of pCF10 (30), which are negative regulatory proteins of pAD1 (45) and pCF10 (16), respectively. All ORFs in the regulatory region were arranged in the same FIG.…”

Section: Resultsmentioning

confidence: 99%

“…Of these plasmids, the pheromone-related conjugation systems best studied are pAD1 (60 kb) (6,7,9,12,23,33,35,36,40,41,44,45) and pCF10 (54 kb) (4, 30, 37), which confer responses to sex pheromones cAD1 and cCF10, respectively. pAD1 encodes a hemolysin-bacteriocin mediated by the same genetic determinant (2, 3, 5, 26) and belongs to incompatibility group incHly (13,25).…”

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Physical mapping of the conjugative bacteriocin plasmid pPD1 of Enterococcus faecalis and identification of the determinant related to the pheromone response

et al. 1995

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The pheromone-responding conjugative bacteriocin plasmid pPD1 (59 kb) of Enterococcus faecalis was mapped physically by using a relational clone approach, and transposon analysis with Tn917 (Em r ) or Tn916 (Tc r ) facilitated the location of the bacteriocin-related genes in a segment of about 6.7 kb. Tn917 insertions within a 3-kb region resulted in constitutive clumping. The nucleotide sequence of the region that included the insertions giving rise to constitutive clumping was determined. The region of pPD1 spanned about 8 kb and was found to contain a number of open reading frames, some of which were named on the basis of homologies with two other pheromone-responding plasmids, pAD1 and pCF10. The genes were arranged in the sequence repB-repA-traC-traB-traA-ipd-traE-traF-orfY-sea-1 with all but repB and traA oriented in the same (left-toright) direction. traC and traB corresponded, respectively, to traC and traB of pAD1 and to prgY and prgZ of pCF10.Certain conjugative plasmids in Enterococcus faecalis transfer at a high frequency in broth mating, a phenomenon relating to a response to specific peptide sex pheromones secreted by potential recipients. The sex pheromone induces the synthesis of a surface aggregation substance that facilitates the formation of a mating aggregate (5-7, 14, 15, 18, 48). When a given plasmid is acquired, secretion of the related pheromone is prevented; however, unrelated pheromones continue to be produced. In addition, the plasmid itself determines the production of a peptide that acts as a competitive inhibitor of the corresponding pheromone (8,10,11,27,32).Several pheromone-responding plasmids have been reported, and the plasmids in E. faecalis have been found to encode such traits as hemolysin-bacteriocin production, UV resistance, and drug resistance (5-7). Of these plasmids, the pheromone-related conjugation systems best studied are pAD1 (60 kb) (6,7,9,12,23,33,35,36,40,41,44,45) and pCF10 (54 kb) (4, 30, 37), which confer responses to sex pheromones cAD1 and cCF10, respectively. pAD1 encodes a hemolysin-bacteriocin mediated by the same genetic determinant (2, 3, 5, 26) and belongs to incompatibility group incHly (13,25). Most (over 90%) of the Hly-Bac plasmids identified in clinical isolates are identical and exhibit extensive homology to pAD1, respond to cAD1, and represent incompatibility group incHly (24,25,29). pAD1 is a representative of this group. pCF10 carries a tetracycline resistance determinant located on transposon Tn925. Genes involved in regulation of the pheromone response have been identified and are clustered in a 7-kb region on each of the plasmids. In pAD1, these genes are arranged in the order traE1-iad-traA-traC-traB (6). The traE1 product is a key positive regulator for expression of downstream structural genes, including determinant asa-1 for the aggregation substance and other conjugation-related genes.The traA product has been shown to represent a key negative regulator of traE1 expression. The traB product is involved in shutdown of endogenous cAD1 p...

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

Physical mapping of the conjugative bacteriocin plasmid pPD1 of Enterococcus faecalis and identification of the determinant related to the pheromone response

et al. 1995

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“…While, for the plasmid transfer itself few data are available, much more is known about the aggregation reaction. In the case of the most extensively studied sex-pheromone plasmid, pAD1, a region of about 12 kb has been sequenced, containing the genes for AS (asal; coding for a protein of 137 kDa; Galli et al, 1990), a surface exclusion protein (seal ; coding for a cell surface protein of 97 kDa that prevents plasmid transfer to cells already containing PAD1 ; Weidlich et al, 1992), a postulated positive regulator (truEl; coding for a peptide of 118 amino acids; Pontius and Clewell, 1992b) processed into peptide iADl that competitively counteracts sex-pheromone CAD1 ; Clewell et al, 1990), a postulated negative regulator (truA, Pontius and Clewell 1992a;see also below) and several other open reading frames (see also Fig. 1C).…”

Section: Sex Pheromone Plasmid Padl-encoded Aggregation Substance Of mentioning

confidence: 99%

Sex pheromone plasmid pAD1‐encoded aggregation substance of Enterococcus faecalis is positively regulated in trans by traE1

Muscholl¹,

Galli²,

Wanner

et al. 1993

European Journal of Biochemistry

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Sex-pheromone-plasmid-bearing strains of Enterococcus faecalis react with sex pheromone to induce the expression of an adhesin, the so-called aggregation substance, on their cell surface. Here we show that, by complementation studies, for sex-pheromone plasmid pADl, expression of the structural gene asal, coding for an aggregation substance, is mediated by a diffusible factor encoded on pAD1. We were able to demonstrate that a small open reading frame, traEl, is sufficient for transcription of the operon containing asal. A model for expression of asal under the influence of the positive regulator is presented, which is supported by our observation that regulation involves an all-or-nothing induction phenomenon, leading to cells either fully expressing asal or not at all.Plasmid-free strains of Enterococcus faecalis excrete short peptides (7 or 8 amino acids in length), the so-called sex pheromones, which can be 'sensed' by cells containing a corresponding sex-pheromone plasmid. These sex-pheromone-plasmid-bearing cells react by expressing a plasmidencoded adhesin, the so-called aggregation substance (AS), on their surface. By interaction of AS with a specific binding substance, which is also present on plasmid-free cells, large macroscopically visible aggregates form with the subsequent and highly efficient transfer of the plasmid. (Conjugative plasmids of E. faecalis not responding to sex pheromones are transferred in broth four orders or five orders of magnitude less efficiently than sex-pheromone plasmids.) After an E. faecalis cell has received a sex-pheromone plasmid, it shuts down the production of the corresponding sex pheromone, whereas sex pheromones specific to other plasmids continue to be produced. By this mechanism, a multitude of different sex-pheromone plasmids can accumulate in a single cell (see Clewell and Weaver 1989; Dunny, 1990). Little is known about the advantage those plasmids offer the cell; our recent data indicate that the adhesin might at least contribute to the virulence of E. fuecalis strains (Kreft et al., 1992).Obviously, the response to sex pheromones is strictly regulated. While, for the plasmid transfer itself few data are available, much more is known about the aggregation reaction. In the case of the most extensively studied sex-pheromone plasmid, pAD1, a region of about 12 kb has been sequenced, containing the genes for AS (asal; coding for a protein of 137 kDa; Galli et al., 1990)

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“…1 (29). The nucleotide sequence for traEl-iad-traA has been reported (5,21,22), as has the traB sequence (1).…”

mentioning

confidence: 99%

“…nucleotide sequence determination, introduction of plasmids into bacteria by conjugation or electroporation, and other aspects of the methodology were essentially as previously described (2,7,21,26).…”

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Characterization of the traC determinant of the Enterococcus faecalis hemolysin-bacteriocin plasmid pAD1: binding of sex pheromone

Tanimoto

Clewell

1993

J Bacteriol

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pAD1, a conjugative, 60-kb, hemolysin-bacteriocin plasmid in Enterococcus faecalis, encodes a mating response to a small peptide sex pheromone, cAD1, secreted by potential recipient bacteria. A gene, traC, encoding a 60.7-kDa protein with a typical amino terminal signal peptide, was identified within a region that appears to encode a product that binds to exogenous pheromone. A cloned segment of DNA containing traC resulted in specific binding of cells to synthetic cAD1. The putative traC product has strong similarity to a product of the E. faecalis plasmid pCF10 as well as oligopeptide binding proteins of Escherichia coli, Salmonella typhimurium, and Bacillus subtilis.

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Regulation of the pAD1-encoded sex pheromone response in Enterococcus faecalis: nucleotide sequence analysis of traA

Cited by 39 publications

References 42 publications

Physical mapping of the conjugative bacteriocin plasmid pPD1 of Enterococcus faecalis and identification of the determinant related to the pheromone response

Physical mapping of the conjugative bacteriocin plasmid pPD1 of Enterococcus faecalis and identification of the determinant related to the pheromone response

Sex pheromone plasmid pAD1‐encoded aggregation substance of Enterococcus faecalis is positively regulated in trans by traE1

Characterization of the traC determinant of the Enterococcus faecalis hemolysin-bacteriocin plasmid pAD1: binding of sex pheromone

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