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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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Expression of qrr1 increases with SypG-mediated activation of syp locus. Levels of qrr1 expression in WT (TIM303), ΔsypK (TIM395), and ΔsypL (KV6629) harboring pKV282 (vector) or pCLD56 (sypG++). The wild-type strain ES114 harboring pCLD56 was used as the nonfluorescent control for quantifying fluorescence levels. Graphical and error bars represent average and standard deviation of three biological replicates, respectively, from a representative experiment performed twice. Comparisons with significance based on two-way ANOVA with Tukey's multiple comparisons test shows significance **(P-value <0.001) between columns, except for the comparison labeled nonsignificant (n.s.) and for comparisons between columns for vector controls, which are also nonsignificant.
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fig08: Expression of qrr1 increases with SypG-mediated activation of syp locus. Levels of qrr1 expression in WT (TIM303), ΔsypK (TIM395), and ΔsypL (KV6629) harboring pKV282 (vector) or pCLD56 (sypG++). The wild-type strain ES114 harboring pCLD56 was used as the nonfluorescent control for quantifying fluorescence levels. Graphical and error bars represent average and standard deviation of three biological replicates, respectively, from a representative experiment performed twice. Comparisons with significance based on two-way ANOVA with Tukey's multiple comparisons test shows significance **(P-value <0.001) between columns, except for the comparison labeled nonsignificant (n.s.) and for comparisons between columns for vector controls, which are also nonsignificant.

Mentions: We thus assayed qrr1 expression in cells harboring either a multicopy plasmid containing sypG (pCLD56) or a vector control (pKV282). Wild-type cells that overexpressed sypG exhibited ˜2.4-fold higher qrr1 expression levels than did vector-containing cells (Fig. 8). This level of qrr1 activity was comparable to that in cells overexpressing sypK (Fig. 4). The level of qrr1 activation in response to sypG decreased to 1.5-fold in the absence of sypK (Fig. 8). In addition, the 2.7-fold qrr1 activation by sypG in a ΔsypL mutant was comparable to the response in wild-type cells, confirming our results that showed qrr1 expression is independent of sypL induction (Fig. 4). Thus, our results show that conditions that lead to transcription of the syp locus, that is, biofilm formation, also activate qrr1 expression.


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)

Expression of qrr1 increases with SypG-mediated activation of syp locus. Levels of qrr1 expression in WT (TIM303), ΔsypK (TIM395), and ΔsypL (KV6629) harboring pKV282 (vector) or pCLD56 (sypG++). The wild-type strain ES114 harboring pCLD56 was used as the nonfluorescent control for quantifying fluorescence levels. Graphical and error bars represent average and standard deviation of three biological replicates, respectively, from a representative experiment performed twice. Comparisons with significance based on two-way ANOVA with Tukey's multiple comparisons test shows significance **(P-value <0.001) between columns, except for the comparison labeled nonsignificant (n.s.) and for comparisons between columns for vector controls, which are also nonsignificant.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig08: Expression of qrr1 increases with SypG-mediated activation of syp locus. Levels of qrr1 expression in WT (TIM303), ΔsypK (TIM395), and ΔsypL (KV6629) harboring pKV282 (vector) or pCLD56 (sypG++). The wild-type strain ES114 harboring pCLD56 was used as the nonfluorescent control for quantifying fluorescence levels. Graphical and error bars represent average and standard deviation of three biological replicates, respectively, from a representative experiment performed twice. Comparisons with significance based on two-way ANOVA with Tukey's multiple comparisons test shows significance **(P-value <0.001) between columns, except for the comparison labeled nonsignificant (n.s.) and for comparisons between columns for vector controls, which are also nonsignificant.
Mentions: We thus assayed qrr1 expression in cells harboring either a multicopy plasmid containing sypG (pCLD56) or a vector control (pKV282). Wild-type cells that overexpressed sypG exhibited ˜2.4-fold higher qrr1 expression levels than did vector-containing cells (Fig. 8). This level of qrr1 activity was comparable to that in cells overexpressing sypK (Fig. 4). The level of qrr1 activation in response to sypG decreased to 1.5-fold in the absence of sypK (Fig. 8). In addition, the 2.7-fold qrr1 activation by sypG in a ΔsypL mutant was comparable to the response in wild-type cells, confirming our results that showed qrr1 expression is independent of sypL induction (Fig. 4). Thus, our results show that conditions that lead to transcription of the syp locus, that is, biofilm formation, also activate qrr1 expression.

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
Related in: MedlinePlus