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The putative oligosaccharide translocase SypK connects biofilm formation with quorum signaling in Vibrio fischeri.

Miyashiro T, Oehlert D, Ray VA, Visick KL, Ruby EG - Microbiologyopen (2014)

Bottom Line: We found that overexpression of sypK, which encodes a putative oligosaccharide translocase, is sufficient to activate qrr1, and, in addition, this effect appears to depend on the kinase activity of the sensor LuxQ.Finally, we found that induction of the syp locus by overexpression of sypG was sufficient to activate qrr1 levels.Together, our results show how conditions that promote biofilm formation impact the quorum-signaling network in V. fischeri, and further highlight the integrated nature of the regulatory circuits involved in complex bacterial behaviors.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Eberly College of Science, The Pennsylvania State University, University Park, Pennsylvania, 16802; Department of Medical Microbiology and Immunology, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, 53706.

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Model of the core quorum-signaling (QS) system in Vibrio fischeri. The outputs of the QS systems AinS/AinR and LuxS/LuxP/LuxQ converge on the LuxU/LuxO phosphorelay. Phosphorylated LuxO activates transcription of the small regulatory RNA Qrr1 that posttranscriptionally represses litR, which encodes the transcription factor LitR. In this study, we show that SypK modulates QS by affecting the kinase activity of LuxQ (indicated by the yellow arrow).
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fig01: Model of the core quorum-signaling (QS) system in Vibrio fischeri. The outputs of the QS systems AinS/AinR and LuxS/LuxP/LuxQ converge on the LuxU/LuxO phosphorelay. Phosphorylated LuxO activates transcription of the small regulatory RNA Qrr1 that posttranscriptionally represses litR, which encodes the transcription factor LitR. In this study, we show that SypK modulates QS by affecting the kinase activity of LuxQ (indicated by the yellow arrow).

Mentions: Vibrio fischeri is a marine bacterium that uses QS to regulate a multitude of cellular processes, including bioluminescence, motility, and colonization of its natural host, the Hawaiian bobtail squid, Euprymna scolopes (Nyholm and McFall-Ngai 2004; Miyashiro and Ruby 2012; Stabb and Visick 2013; Verma and Miyashiro 2013). The LuxR-LuxI QS system directly regulates the lux genes, which encode the light-producing enzyme luciferase and several proteins involved in light production and other activities. LuxR is a transcription factor activated by the autoinducer N-3-oxohexanoyl-homoserine lactone (3-oxo-C6), which is produced by the synthase LuxI. V. fischeri possesses additional QS systems that converge on a signaling cascade that, unlike the LuxR-LuxI system, is conserved among all Vibrionaceae members (Milton 2006). At its core is a phosphorelay composed of the histidine phosphotransfer protein LuxU and the response regulator LuxO (Fig. 1). Based primarily on the studies of the analogous phosphorelay in Vibrio harveyi, LuxU is predicted to become phosphorylated on a conserved histidine residue by the kinases AinR and LuxQ under conditions of low autoinducer concentrations, for example, low cell density (Freeman and Bassler 1999a, 1999b; Ray and Visick 2012). Whereas AinR appears to serve as the receptor for the AinS-derived autoinducer N-octonoyl-homoserine lactone (C8) (Gilson et al. 1995; Kimbrough and Stabb 2013), the periplasmic protein LuxP is thought, based on work in V. harveyi, to bind to the furanosyl borate diester, autoinducer-2 (AI-2), which modulates the kinase activity of LuxQ toward LuxU (Neiditch et al. 2005, 2006). Upon phosphorylation, LuxU is predicted to donate the phosphoryl group to a conserved aspartic acid residue of LuxO, which can then activate transcription of qrr1 (Miyashiro et al. 2010). The RNA chaperone Hfq assists the small regulatory RNA (sRNA) Qrr1 in the posttranscriptional repression of LitR, a global transcription factor that regulates motility, host colonization factors, and bioluminescence (Fidopiastis et al. 2002; Miyashiro et al. 2010; Cao et al. 2012). The net effect of the integrated QS systems is that under high cell density (i.e., in the presence of autoinducers) LuxO becomes de-phosphorylated, which leads to low qrr1 expression and the ability of V. fischeri to fully activate the lux genes.


The putative oligosaccharide translocase SypK connects biofilm formation with quorum signaling in Vibrio fischeri.

Miyashiro T, Oehlert D, Ray VA, Visick KL, Ruby EG - Microbiologyopen (2014)

Model of the core quorum-signaling (QS) system in Vibrio fischeri. The outputs of the QS systems AinS/AinR and LuxS/LuxP/LuxQ converge on the LuxU/LuxO phosphorelay. Phosphorylated LuxO activates transcription of the small regulatory RNA Qrr1 that posttranscriptionally represses litR, which encodes the transcription factor LitR. In this study, we show that SypK modulates QS by affecting the kinase activity of LuxQ (indicated by the yellow arrow).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263508&req=5

fig01: Model of the core quorum-signaling (QS) system in Vibrio fischeri. The outputs of the QS systems AinS/AinR and LuxS/LuxP/LuxQ converge on the LuxU/LuxO phosphorelay. Phosphorylated LuxO activates transcription of the small regulatory RNA Qrr1 that posttranscriptionally represses litR, which encodes the transcription factor LitR. In this study, we show that SypK modulates QS by affecting the kinase activity of LuxQ (indicated by the yellow arrow).
Mentions: Vibrio fischeri is a marine bacterium that uses QS to regulate a multitude of cellular processes, including bioluminescence, motility, and colonization of its natural host, the Hawaiian bobtail squid, Euprymna scolopes (Nyholm and McFall-Ngai 2004; Miyashiro and Ruby 2012; Stabb and Visick 2013; Verma and Miyashiro 2013). The LuxR-LuxI QS system directly regulates the lux genes, which encode the light-producing enzyme luciferase and several proteins involved in light production and other activities. LuxR is a transcription factor activated by the autoinducer N-3-oxohexanoyl-homoserine lactone (3-oxo-C6), which is produced by the synthase LuxI. V. fischeri possesses additional QS systems that converge on a signaling cascade that, unlike the LuxR-LuxI system, is conserved among all Vibrionaceae members (Milton 2006). At its core is a phosphorelay composed of the histidine phosphotransfer protein LuxU and the response regulator LuxO (Fig. 1). Based primarily on the studies of the analogous phosphorelay in Vibrio harveyi, LuxU is predicted to become phosphorylated on a conserved histidine residue by the kinases AinR and LuxQ under conditions of low autoinducer concentrations, for example, low cell density (Freeman and Bassler 1999a, 1999b; Ray and Visick 2012). Whereas AinR appears to serve as the receptor for the AinS-derived autoinducer N-octonoyl-homoserine lactone (C8) (Gilson et al. 1995; Kimbrough and Stabb 2013), the periplasmic protein LuxP is thought, based on work in V. harveyi, to bind to the furanosyl borate diester, autoinducer-2 (AI-2), which modulates the kinase activity of LuxQ toward LuxU (Neiditch et al. 2005, 2006). Upon phosphorylation, LuxU is predicted to donate the phosphoryl group to a conserved aspartic acid residue of LuxO, which can then activate transcription of qrr1 (Miyashiro et al. 2010). The RNA chaperone Hfq assists the small regulatory RNA (sRNA) Qrr1 in the posttranscriptional repression of LitR, a global transcription factor that regulates motility, host colonization factors, and bioluminescence (Fidopiastis et al. 2002; Miyashiro et al. 2010; Cao et al. 2012). The net effect of the integrated QS systems is that under high cell density (i.e., in the presence of autoinducers) LuxO becomes de-phosphorylated, which leads to low qrr1 expression and the ability of V. fischeri to fully activate the lux genes.

Bottom Line: We found that overexpression of sypK, which encodes a putative oligosaccharide translocase, is sufficient to activate qrr1, and, in addition, this effect appears to depend on the kinase activity of the sensor LuxQ.Finally, we found that induction of the syp locus by overexpression of sypG was sufficient to activate qrr1 levels.Together, our results show how conditions that promote biofilm formation impact the quorum-signaling network in V. fischeri, and further highlight the integrated nature of the regulatory circuits involved in complex bacterial behaviors.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Eberly College of Science, The Pennsylvania State University, University Park, Pennsylvania, 16802; Department of Medical Microbiology and Immunology, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, 53706.

Show MeSH