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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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Overexpression of sypK enhances qrr1-dependent motility. (A) Soft-agar motility migration distances by WT (ES114; closed symbols) and Δqrr1 (TIM305; open symbols) harboring vector (pTM214; circles) or Ptrc-sypK (pTM367; squares). TBSW motility plates contained chloramphenicol and 100 μmol/L IPTG. Points and error bars (too small to visualize) represent means and standard deviations of spot diameter (in mm) for quadruplicate biological replicates, respectively. Experiment was performed twice, with similar results. (B) Image of motility plate used in (A) at 4 h.
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fig05: Overexpression of sypK enhances qrr1-dependent motility. (A) Soft-agar motility migration distances by WT (ES114; closed symbols) and Δqrr1 (TIM305; open symbols) harboring vector (pTM214; circles) or Ptrc-sypK (pTM367; squares). TBSW motility plates contained chloramphenicol and 100 μmol/L IPTG. Points and error bars (too small to visualize) represent means and standard deviations of spot diameter (in mm) for quadruplicate biological replicates, respectively. Experiment was performed twice, with similar results. (B) Image of motility plate used in (A) at 4 h.

Mentions: Previous studies have shown that V. fischeri also uses the LuxU-LuxO QS pathway to regulate motility (Lupp and Ruby 2005; Cao et al. 2012). In particular, V. fischeri cells become hypermotile in the absence of litR. To test whether SypK has an effect on motility, we examined on soft agar the motility of cells expressing sypK. Compared to wild-type motility levels, cells with induced sypK expression were hypermotile (Fig. 5A and B). Furthermore, this hypermotility phenotype depended on Qrr1, as shown by the overall reduced motility associated with mutants containing the Δqrr1 allele. Together, these results demonstrate that expression of sypK leads to higher motility due to increased qrr1 transcription.


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)

Overexpression of sypK enhances qrr1-dependent motility. (A) Soft-agar motility migration distances by WT (ES114; closed symbols) and Δqrr1 (TIM305; open symbols) harboring vector (pTM214; circles) or Ptrc-sypK (pTM367; squares). TBSW motility plates contained chloramphenicol and 100 μmol/L IPTG. Points and error bars (too small to visualize) represent means and standard deviations of spot diameter (in mm) for quadruplicate biological replicates, respectively. Experiment was performed twice, with similar results. (B) Image of motility plate used in (A) at 4 h.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig05: Overexpression of sypK enhances qrr1-dependent motility. (A) Soft-agar motility migration distances by WT (ES114; closed symbols) and Δqrr1 (TIM305; open symbols) harboring vector (pTM214; circles) or Ptrc-sypK (pTM367; squares). TBSW motility plates contained chloramphenicol and 100 μmol/L IPTG. Points and error bars (too small to visualize) represent means and standard deviations of spot diameter (in mm) for quadruplicate biological replicates, respectively. Experiment was performed twice, with similar results. (B) Image of motility plate used in (A) at 4 h.
Mentions: Previous studies have shown that V. fischeri also uses the LuxU-LuxO QS pathway to regulate motility (Lupp and Ruby 2005; Cao et al. 2012). In particular, V. fischeri cells become hypermotile in the absence of litR. To test whether SypK has an effect on motility, we examined on soft agar the motility of cells expressing sypK. Compared to wild-type motility levels, cells with induced sypK expression were hypermotile (Fig. 5A and B). Furthermore, this hypermotility phenotype depended on Qrr1, as shown by the overall reduced motility associated with mutants containing the Δqrr1 allele. Together, these results demonstrate that expression of sypK leads to higher motility due to increased qrr1 transcription.

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