Analysis of expression of prgX , a key negative regulator of the transfer of the Enterococcus faecalis pheromone-inducible plasmid pCF10 1 1Edited by M. Gottesman

Bae, Taeok; Clerc-Bardin, Sylvie; Dunny, Gary M.

doi:10.1006/jmbi.2000.3628

Cited by 47 publications

(79 citation statements)

References 35 publications

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“…Purification of total RNA and Northern hybridization analysis were performed as described in ref. 23 except that cells were grown in M9-YE overnight (15 hr), diluted 1:5, and grown for an additional 20 min. RNA was isolated by using the BIO101-FastRNA RE kit (Qbiogene), and RNA samples were separated on a 2% agarose gel containing 2% formaldehyde and 1ϫ Mops (20 mM morpholinopropanesulfonic acid).…”

Section: Methodsmentioning

confidence: 99%

A paracrine peptide sex pheromone also acts as an autocrine signal to induce plasmid transfer and virulence factor expressionin vivo

Chandler

Hirt

Dunny

2005

Proc. Natl. Acad. Sci. U.S.A.

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The peptide pheromone cCF10 of Enterococcus faecalis is an intercellular signal for induction of conjugative transfer of plasmid pCF10 from donor cells to recipient cells. When a donor cell is exposed to recipient-produced cCF10, expression of the pCF10-encoded aggregation substance of pCF10 (Asc10) and other conjugation gene products is activated. Asc10 also increases enterococcal virulence in several models, and when donor cells are grown in animals or in plasma, Asc10 expression is induced by means of the cCF10-sensing machinery. Plasmid pCF10 carries two genes that function to prevent self-induction by endogenous cCF10 in donor cells. The membrane protein PrgY reduces endogenous pheromone activity in donor cells, and the inhibitor peptide iCF10 neutralizes the residual endogenous cCF10 that escapes PrgY. In the current study, we found that E. faecalis strains with allelic replacements abolishing active cCF10 production showed reduced ability to acquire pCF10 by conjugation; prgY-null mutations had no phenotype in the cCF10-negative strains. We observed that expression of the mRNA for iCF10 was reduced in this background and that these mutations also blocked plasma induction of Asc10 expression. These findings support a model in which plasma induction in wild-type donors results from iCF10 inactivation by a plasma component, causing disruption of a precisely maintained balance of iCF10 to cCF10 activity and allowing subsequent induction by endogenous cCF10. Although cCF10 has traditionally been viewed as an intercellular signal, these results show that pCF10 has also adapted cCF10 as an autocrine signal that activates expression of virulence and conjugation functions.cell-cell signaling ͉ Enterococcus ͉ horizontal gene transfer ͉ pCF10

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Section: Methodsmentioning

confidence: 99%

A paracrine peptide sex pheromone also acts as an autocrine signal to induce plasmid transfer and virulence factor expressionin vivo

Chandler

Hirt

Dunny

2005

Proc. Natl. Acad. Sci. U.S.A.

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“…Once taken up by donor cells, I competes with C for binding to PrgX; the two peptides bind to the same subdomain of PrgX but have opposing effects on PrgX structure and function (11). It has been suggested that in the absence of recipient cells, the basal level expression of I keeps the expression of conjugation genes OFF in donor cells (11,12). When recipient cells are in close proximity, the increased C level overcomes the inhibition of I, thus allowing donor cells to mount a mating response.…”

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confidence: 99%

Antagonistic self-sensing and mate-sensing signaling controls antibiotic-resistance transfer

Chatterjee

Cook

Shu

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Conjugation is one of the most common ways bacteria acquire antibiotic resistance, contributing to the emergence of multidrugresistant "superbugs." Bacteria of the genus Enterococcus faecalis are highly antibiotic-resistant nosocomial pathogens that use the mechanism of conjugation to spread antibiotic resistance between resistance-bearing donor cells and resistance-deficient recipient cells. Here, we report a unique quorum sensing-based communication system that uses two antagonistic signaling molecules to regulate conjugative transfer of tetracycline-resistance plasmid pCF10 in E. faecalis. A "mate-sensing" peptide sex pheromone produced by recipient cells is detected by donor cells to induce conjugative genetic transfer. Using mathematical modeling and experimentation, we show that a second antagonistic "self-sensing" signaling peptide, previously known to suppress self-induction of donor cells, also serves as a classic quorum-sensing signal for donors that functions to reduce antibiotic-resistance transfer at high donor density. This unique form of quorum sensing may provide a means of limiting the spread of the plasmid and present opportunities to control antibiotic-resistance transfer through manipulation of intercellular signaling, with implications in the clinical setting.

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“…Both regulatory molecules are produced from one transcript that initiates at the Qa promoter (P Qa , Fig. 1B) (Bae et al, 2000). A 5′ fragment of the message comprises the 102 nt Qa (antisense to prgQ) RNA (Fig.…”

Section: The Pheromone-sensitive Switch and Initiation Of A Mating Rementioning

confidence: 99%

“…A 5′ fragment of the message comprises the 102 nt Qa (antisense to prgQ) RNA (Fig. 1B) (Bae et al, 2000. A direct interaction between the abundant prgQ transcript Qs and Qa is believed to prevent transcripts initiating at P Q from reaching downstream genes, thereby providing a roadblock to transcription beyond the prgQ gene .…”

Section: The Pheromone-sensitive Switch and Initiation Of A Mating Rementioning

confidence: 99%

“…PrgX protein was known to be required for proper post-transcriptional processing of the Qa-prgX transcript to produce Qa RNA . Additionally, PrgX is an essential positive regulator of its own expression, although it does not act on its own promoter (P Qa ) (Bae et al, 2000). Interestingly, PrgX was found to require the DNA loop in order to regulate its own production and that of Qa RNA; indicating that the DNA loop regulates transcription from both strands of pCF10 DNA.…”

Section: The Pheromone-sensitive Switch and Initiation Of A Mating Rementioning

confidence: 99%

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Pheromone-inducible conjugation in Enterococcus faecalis: A model for the evolution of biological complexity?

Kozlowicz

Dworkin

Dunny

2006

International Journal of Medical Microbiology

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Pheromone-inducible transfer of the plasmid pCF10 in Enterococcus faecalis is regulated using a complicated network of proteins and RNAs. The plasmid itself has been assembled from parts garnered from a variety of sources, and many aspects of the system resemble a biological kluge. Recently several new functions of various pCF10 gene products that participate in regulation of plasmid transfer have been identified. The results indicate that selective pressures controlling the evolution of the plasmid have produced a highly complex regulatory network with multiple biological functions that may serve well as a model for the evolution of biological complexity.

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Analysis of expression of prgX , a key negative regulator of the transfer of the Enterococcus faecalis pheromone-inducible plasmid pCF10 1 1Edited by M. Gottesman

Cited by 47 publications

References 35 publications

A paracrine peptide sex pheromone also acts as an autocrine signal to induce plasmid transfer and virulence factor expressionin vivo

A paracrine peptide sex pheromone also acts as an autocrine signal to induce plasmid transfer and virulence factor expressionin vivo

Antagonistic self-sensing and mate-sensing signaling controls antibiotic-resistance transfer

Pheromone-inducible conjugation in Enterococcus faecalis: A model for the evolution of biological complexity?

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