Structural Basis of the Sensor-Synthase Interaction in Autoinduction of the Quorum Sensing Signal DSF Biosynthesis

Chen, Zhihong; He, Yanan; Lim, Siew Choo; Qamra, Rohini; Walsh, Martin A.; Zhang, Lian-Hui; Song, Haiwei

doi:10.1016/j.str.2010.06.011

Cited by 45 publications

(76 citation statements)

References 44 publications

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“…In X. campestris pv. campestris, RpfC physically interacts with the RpfF active site, inhibiting DSF synthesis activity (12,22,24). RpfC has also been shown to repress the RpfF activity of X. fastidiosa (25).…”

Section: Discussionmentioning

confidence: 99%

“…It has also been suggested that in X. campestris pv. campestris, RpfC could play a positive-feedback role in DSF synthesis, liberating active RpfF upon the detection of DSF molecules (22). Assuming similar mechanisms in S. maltophilia, we hypothesize that DSF production in RpfC-RpfF-1 strains is due to the presence of a competent sensor input domain, i.e., composed of 10 TM regions, in RpfC-1, which would enable the liberation of active RpfF-1 upon DSF detection and the subsequent synthesis of DSF.…”

Section: Discussionmentioning

confidence: 99%

“…However, the residues that form the substrate binding pocket (Leu136, Gly137, Gly138, Gly85, Leu276, Met170, and Trp258), as well as those involved in catalysis (Glu141, Glu161), in X. campestris pv. campestris (22) are conserved in the two variants (Fig. 2B).…”

Section: S Maltophilia Harbors Twomentioning

confidence: 94%

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Two Different rpf Clusters Distributed among a Population of Stenotrophomonas maltophilia Clinical Strains Display Differential Diffusible Signal Factor Production and Virulence Regulation

et al. 2014

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The quorum-sensing (QS) system present in the emerging nosocomial pathogen Stenotrophomonas maltophilia is based on the signaling molecule diffusible signal factor (DSF). Production and detection of DSF are governed by the rpf cluster, which encodes the synthase RpfF and the sensor RpfC, among other components. Despite a well-studied system, little is known about its implication in virulence regulation in S. maltophilia. Here, we have analyzed the rpfF gene from 82 S. maltophilia clinical isolates. Although rpfF was found to be present in all of the strains, it showed substantial variation, with two populations (rpfF-1 and rpfF-2) clearly distinguishable by the N-terminal region of the protein. Analysis of rpfC in seven complete genome sequences revealed a corresponding variability in the N-terminal transmembrane domain of its product, suggesting that each RpfF variant has an associated RpfC variant. We show that only RpfC-RpfF-1 variant strains display detectable DSF production. Heterologous rpfF complementation of ⌬rpfF mutants of a representative strain of each variant suggests that RpfF-2 is, however, functional and that the observed DSF-deficient phenotype of RpfC-RpfF-2 variant strains is due to permanent repression of RpfF-2 by RpfC-2. This is corroborated by the ⌬rpfC mutant of the RpfC-RpfF-2 representative strain. In line with this observations, deletion of rpfF from the RpfC-RpfF-1 strain leads to an increase in biofilm formation, a decrease in swarming motility, and relative attenuation in the Caenorhabditis elegans and zebrafish infection models, whereas deletion of the same gene from the representative RpfC-RpfF-2 strain has no significant effect on these virulence-related phenotypes. Q uorum sensing (QS) is a bacterial cell-cell communication process that allows bacteria to synchronize particular behaviors on a population-wide scale. Within current knowledge, QS in Stenotrophomonas maltophilia depends on the diffusible signal factor QS (DSF-QS) system, which is based mainly on the fatty acid DSF (cis-11-methyl-2-dodecenoic acid) (1, 2). DSF synthesis is fully dependent on RpfF, an enoyl coenzyme A hydratase encoded by the rpf (regulation of pathogenicity factors) cluster, a set of genes that includes all of the components necessary for the synthesis and detection of DSF molecules. In addition to RpfF, rpf encodes the aconitase RpfA, the fatty acid ligase RpfB, the two-component sensor-effector hybrid system RpfC, and the cytoplasmic regulator element RpfG (1, 2). The DSF-QS system was first described in the phytopathogen Xanthomonas campestris pv. campestris, where it plays an important role in virulence regulation (3). Since then, this system has been described in several members of the order Xanthomonadales, including the genera Xanthomonas, Xylella, and Stenotrophomonas, as well as in members of the order Burkholderiales (1, 3-5). The specific functions regulated by the DSF-QS system are dependent on the species, but it has been suggested that it controls several virulence-related phenotypes (6...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Two Different rpf Clusters Distributed among a Population of Stenotrophomonas maltophilia Clinical Strains Display Differential Diffusible Signal Factor Production and Virulence Regulation

et al. 2014

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“…This was surprising since in Xanthomonas campestris, the phenotypes of a ⌬rpfF strain could be rescued by the addition of DSF to the growth medium (5,15), and current models of DSF-mediated signaling involving physical interactions of RpfF and RpfC (16,21) do not account for such an observation. In X. campestris, RpfF has been demonstrated to function only as a DSF synthase whose catalytic activity is repressed by physical interactions with RpfC (16,21) and is apparently derepressed upon the interaction of DSF with RpfC (21). RpfF has not been suggested to aid RpfC in the signaling process.…”

mentioning

confidence: 98%

Diffusible Signal Factor (DSF) Synthase RpfF of Xylella fastidiosa Is a Multifunction Protein Also Required for Response to DSF

Ionescu

Baccari

Silva

et al. 2013

Journal of Bacteriology

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c Xylella fastidiosa, like related Xanthomonas species, employs an Rpf cell-cell communication system consisting of a diffusible signal factor (DSF) synthase, RpfF, and a DSF sensor, RpfC, to coordinate expression of virulence genes. While phenotypes of a ⌬rpfF strain in Xanthomonas campestris could be complemented by its own DSF, the DSF produced by X. fastidiosa (XfDSF) did not restore expression of the XfDSF-dependent genes hxfA and hxfB to a ⌬rpfF strain of X. fastidiosa, suggesting that RpfF is involved in XfDSF sensing or XfDSF-dependent signaling. To test this conjecture, rpfC and rpfF of X. campestris were replaced by those of X. fastidiosa, and the contribution of each gene to the induction of a X. campestris DSF-dependent gene was assessed. As in X. fastidiosa, XfDSF-dependent signaling required both X. fastidiosa proteins RpfF and RpfC. RpfF repressed RpfC signaling activity, which in turn was derepressed by XfDSF. A mutated X. fastidiosa RpfF protein with two substitutions of glutamate to alanine in its active site was incapable of XfDSF production yet enabled a response to XfDSF, indicating that XfDSF production and the response to XfDSF are two separate functions in which RpfF is involved. This mutant was also hypervirulent to grape, demonstrating the antivirulence effects of XfDSF itself in X. fastidiosa. The Rpf system of X. fastidiosa is thus a novel example of a quorum-sensing signal synthase that is also involved in the response to the signal molecule that it synthesizes.

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“…7B). These catalytic Glu residues are also conserved in RpfF of X. campestris, and mutation of either Glu residue abolished DSF synthesis (46 residues for enoyl-CoA hydratase activity are included among those predicted to be involved in the active site (Fig. 7A).…”

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confidence: 99%

The Putative Enoyl-Coenzyme A Hydratase DspI Is Required for Production of the Pseudomonas aeruginosa Biofilm Dispersion Autoinducer cis -2-Decenoic Acid

2013

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In the present study, we report the identification of a putative enoyl-coenzyme A (CoA) hydratase/isomerase that is required for synthesis of the biofilm dispersion autoinducer cis-2-decenoic acid in the human pathogen Pseudomonas aeruginosa. The protein is encoded by PA14_54640 (PA0745), named dspI for dispersion inducer. The gene sequence for this protein shows significant homology to RpfF in Xanthomonas campestris. Inactivation of dspI was shown to abolish biofilm dispersion autoinduction in continuous cultures of P. aeruginosa and resulted in biofilms that were significantly greater in thickness and biomass than those of the parental wild-type strain. Dispersion was shown to be inducible in dspI mutants by the exogenous addition of synthetic cis-2-decenoic acid or by complementation of ⌬dspI in trans under the control of an arabinose-inducible promoter. Mutation of dspI was also shown to abolish cis-2-decenoic acid production, as revealed by gas chromatography-mass spectrometry (GC-MS) analysis of cell-free spent culture medium. The transcript abundance of dspI correlated with cell density, as determined by quantitative reverse transcriptase (RT) PCR. This regulation is consistent with the characterization of cis-2-decenoic acid as a cell-to-cell communication molecule that regulates biofilm dispersion in a cell density-dependent manner. Biofilm dispersion is the terminal stage of the biofilm developmental cycle, where bacteria regulate their escape from a biofilm and transition to a mobile planktonic lifestyle (1, 2). Induction of biofilm dispersion in Pseudomonas aeruginosa occurs naturally when biofilm microcolonies attain a critical size, releasing bacteria as free cells into the surrounding environment (2, 3). Recently, we have reported that the small messenger fatty acid molecule cis-2-decenoic acid (cis-DA), produced by P. aeruginosa in batch and continuous cultures, acts as the autoinducer of biofilm dispersion for P. aeruginosa (3). This molecule has also been shown to induce biofilm dispersion in a range of Gram-negative and Gram-positive bacteria and in the fungal pathogen Candida albicans (3).The autoinducer cis-DA is a fatty acid cell-to-cell communication molecule with structural homology to cis-11-methyl-2-dodecenoic acid (DSF), isolated from Xanthomonas campestris (4, 5). Analogs of DSF have been identified in Burkholderia cenocepacia (cis-2-dodecenoic acid [BDSF]), Streptococcus mutans (trans-2-decenoic acid [SDSF]), and Xylella fastidiosa (trans-2-tetradecenoic acid [XyDSF]) (6-8). Additional, structurally related fatty acid signals have been identified in the genera Burkholderia, Xanthomonas, and Stenotrophomonas (6,[8][9][10][11][12][13]. Fatty acid signals have been shown to regulate a wide range of bacterial behaviors, including virulence, motility, biofilm development, and dispersion (4, 8-10, 12, 14-23).The mechanism of fatty acid signal biosynthesis appears to be widely conserved. DSF biosynthesis in X. campestris is dependent on the gene rpfF, which encodes a putative enoyl-coenzym...

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Structural Basis of the Sensor-Synthase Interaction in Autoinduction of the Quorum Sensing Signal DSF Biosynthesis

Cited by 45 publications

References 44 publications

Two Different rpf Clusters Distributed among a Population of Stenotrophomonas maltophilia Clinical Strains Display Differential Diffusible Signal Factor Production and Virulence Regulation

Two Different rpf Clusters Distributed among a Population of Stenotrophomonas maltophilia Clinical Strains Display Differential Diffusible Signal Factor Production and Virulence Regulation

Diffusible Signal Factor (DSF) Synthase RpfF of Xylella fastidiosa Is a Multifunction Protein Also Required for Response to DSF

The Putative Enoyl-Coenzyme A Hydratase DspI Is Required for Production of the Pseudomonas aeruginosa Biofilm Dispersion Autoinducer cis -2-Decenoic Acid

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