Functional Analysis of TraA, the Sex Pheromone Receptor Encoded by pPD1, in a Promoter Region Essential for the Mating Response in
            <i>Enterococcus faecalis</i>

Horii, Takaaki; Nagasawa, Hiromichi; Nakayama, Jiro

doi:10.1128/jb.184.22.6343-6350.2002

Cited by 11 publications

(13 citation statements)

References 39 publications

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“…The pheromone is then internalized, making use of a host-encoded peptide transport system (for a recent detailed review, see reference 44). A functional analysis of TraA, the intracellular sex pheromone receptor encoded by pPD1, was recently performed (99). When cPD1 is taken up by a pPD1 donor cell, it binds to an intracellular receptor, TraA.…”

Section: Sex Pheromone Plasmidsmentioning

confidence: 99%

Conjugative Plasmid Transfer in Gram-Positive Bacteria

Grohmann

Muth

Espinosa

2003

Microbiol Mol Biol Rev

507

447

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Conjugative transfer of bacterial plasmids is the most efficient way of horizontal gene spread, and it is therefore considered one of the major reasons for the increase in the number of bacteria exhibiting multiple-antibiotic resistance. Thus, conjugation and spread of antibiotic resistance represents a severe problem in antibiotic treatment, especially of immunosuppressed patients and in intensive care units. While conjugation in gram-negative bacteria has been studied in great detail over the last decades, the transfer mechanisms of antibiotic resistance plasmids in gram-positive bacteria remained obscure. In the last few years, the entire nucleotide sequences of several large conjugative plasmids from gram-positive bacteria have been determined. Sequence analyses and data bank comparisons of their putative transfer (tra) regions have revealed significant similarities to tra regions of plasmids from gram-negative bacteria with regard to the respective DNA relaxases and their targets, the origins of transfer (oriT), and putative nucleoside triphosphatases NTP-ases with homologies to type IV secretion systems. In contrast, a single gene encoding a septal DNA translocator protein is involved in plasmid transfer between micelle-forming streptomycetes. Based on these clues, we propose the existence of two fundamentally different plasmid-mediated conjugative mechanisms in gram-positive microorganisms, namely, the mechanism taking place in unicellular gram-positive bacteria, which is functionally similar to that in gram-negative bacteria, and a second type that occurs in multicellular gram-positive bacteria, which seems to be characterized by double-stranded DNA transfer

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Section: Sex Pheromone Plasmidsmentioning

confidence: 99%

Conjugative Plasmid Transfer in Gram-Positive Bacteria

Grohmann

Muth

Espinosa

2003

Microbiol Mol Biol Rev

507

447

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“…faecalis sex-pheromone-responding conjugative plasmids pCF10, pAD1 and pPD1 (Bae & Dunny, 2001;Bae et al, 2002Bae et al, , 2004Horii et al, 2002; for reviews see Clewell & Dunny, 2002;Chandler & Dunny, 2004).…”

Section: Introductionmentioning

confidence: 99%

The TraA relaxase autoregulates the putative type IV secretion-like system encoded by the broad-host-range Streptococcus agalactiae plasmid pIP501

et al. 2006

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The conjugative multiple antibiotic resistance plasmid pIP501 can be transferred and stably maintained in a variety of Gram-positive genera, including multicellular Streptomyces lividans, as well as in Gram-negative Escherichia coli. The 15 putative pIP501 transfer (tra) genes are organized in an operon-like structure terminating in a strong transcriptional terminator. This paper reports co-transcription of the pIP501 tra genes in exponentially growing Enterococcus faecalis JH2-2 cells, as shown by RT-PCR. The tra genes are expressed throughout the life cycle of Ent. faecalis, and the expression level is independent of the growth phase. Electrophoretic mobility shift assays indicated that the TraA relaxase, the first gene of the tra operon, binds to the tra promoter P tra , which partially overlaps with the origin of transfer (oriT ). DNase I footprinting experiments further delimited the TraA binding region and defined the nucleotides bound by TraA. b-Galactosidase assays with P tra -lacZ fusions proved P tra promoter activity, which was strongly repressed when TraA was supplied in trans. Thus, it is concluded that the pIP501 tra operon is negatively autoregulated at the transcriptional level by the conjugative DNA relaxase TraA.

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“…The mismatch between the middle peak and the expected position of tab2 can be due to a non-symmetric binding of TraA at this site or to a misinterpretation of previously published footprinting data. 10 What is surprising, however, is that the frequency of TraA binding to a specific tab site changes under the different conditions used. Namely, TraA alone binds tab1, tab2 and tab3 with probability values of 0.57, 0.17 and 0.26, respectively; TraA-cPD1 binds tab1, tab2 and tab3 with probability values of 0.32, 0.23 and 0.45, respectively; and TraA-iPD1 binds tab1, tab2 and tab3 with probability values of 0.52, 0.22 and 0.26, respectively.…”

Section: Resultsmentioning

confidence: 94%

“…9 More recently, it has been demonstrated by DNA footprinting experiments that TraA specifically recognizes three distinct sites of approximately 26 bp, one strong (tab1) and two weak (tab2 and tab3), located in the DNA region encompassing the ipd and traA genes. 10 It has also been shown by means of Northern blotting analysis that the transcription is up-regulated and down-regulated for the ipd and traA genes, respectively, by exogenous cPD1. 10 In this study, we employed several techniques to analyze the interaction between TraA and specific regions of the pPD1 plasmid both in the presence and in the absence of cPD1 and iPD1 regulator peptides.…”

Section: Introductionmentioning

confidence: 98%

“…10 It has also been shown by means of Northern blotting analysis that the transcription is up-regulated and down-regulated for the ipd and traA genes, respectively, by exogenous cPD1. 10 In this study, we employed several techniques to analyze the interaction between TraA and specific regions of the pPD1 plasmid both in the presence and in the absence of cPD1 and iPD1 regulator peptides. The results show that in the absence of either peptide or in the presence of the inhibitor peptide iPD1, TraA preferentially binds to tab1 and significantly represses ipd activity.…”

Section: Introductionmentioning

confidence: 98%

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