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Oxygen promotes biofilm formation of Shewanella putrefaciens CN32 through a diguanylate cyclase and an adhesin.

Wu C, Cheng YY, Yin H, Song XN, Li WW, Zhou XX, Zhao LP, Tian LJ, Han JC, Yu HQ - Sci Rep (2013)

Bottom Line: DosD regulates the transcription of bpfA operon which encodes seven proteins including a large repetitive adhesin BpfA and its cognate type I secretion system (TISS).Regulation of DosD in aerobic biofilms is heavily dependent on an adhesin BpfA and the TISS.This study offers an insight into the molecular mechanism of oxygen-stimulated biofilm formation by S. putrefaciens CN32.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, University of Science & Technology of China, Hefei, China.

ABSTRACT
Although oxygen has been reported to regulate biofilm formation by several Shewanella species, the exact regulatory mechanism mostly remains unclear. Here, we identify a direct oxygen-sensing diguanylate cyclase (DosD) and reveal its regulatory role in biofilm formation by Shewanella putrefaciens CN32 under aerobic conditions. In vitro and in vivo analyses revealed that the activity of DosD culminates to synthesis of cyclic diguanylate (c-di-GMP) in the presence of oxygen. DosD regulates the transcription of bpfA operon which encodes seven proteins including a large repetitive adhesin BpfA and its cognate type I secretion system (TISS). Regulation of DosD in aerobic biofilms is heavily dependent on an adhesin BpfA and the TISS. This study offers an insight into the molecular mechanism of oxygen-stimulated biofilm formation by S. putrefaciens CN32.

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DosD positively regulates aerobic biofilm production in S. putrefaciens CN32.(A) Schematic of the genetic arrangement of DosD and its domain architecture. (B) Aerobic biofilm mass profiles of WT (white columns) and ΔdosD (gray columns) with simultaneous determination of growth of WT (solid line) and ΔdosD (dashed line). (C) Biofilm formation by ΔdosD complemented by vector-bearing dosD driven by its native promoter. The empty vector was transformed into WT and ΔdosD mutants as a control. (D) Attachment abilities of WT and ΔdosD. The average number of cells attached to the cover slips was counted in randomly picked areas of ten microscopic images. Each experiment was repeated at least three times and representative results are provided. Error bars refer to the standard deviation of the three samples.
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f1: DosD positively regulates aerobic biofilm production in S. putrefaciens CN32.(A) Schematic of the genetic arrangement of DosD and its domain architecture. (B) Aerobic biofilm mass profiles of WT (white columns) and ΔdosD (gray columns) with simultaneous determination of growth of WT (solid line) and ΔdosD (dashed line). (C) Biofilm formation by ΔdosD complemented by vector-bearing dosD driven by its native promoter. The empty vector was transformed into WT and ΔdosD mutants as a control. (D) Attachment abilities of WT and ΔdosD. The average number of cells attached to the cover slips was counted in randomly picked areas of ten microscopic images. Each experiment was repeated at least three times and representative results are provided. Error bars refer to the standard deviation of the three samples.

Mentions: Genome annotation predicts that Sputcn32_3244 in S. putrefaciens CN32 encodes a putative diguanylate cyclase (DGC), composed of an N-terminal globin domain and a C-terminal GGDEF domain (Figure 1A). Given the typical function of the globin domain as a gas sensor16, we therefore postulated that Sputcn32_3244 might encode an oxygen sensing DGC (dosD) in S. putrefaciens CN32.


Oxygen promotes biofilm formation of Shewanella putrefaciens CN32 through a diguanylate cyclase and an adhesin.

Wu C, Cheng YY, Yin H, Song XN, Li WW, Zhou XX, Zhao LP, Tian LJ, Han JC, Yu HQ - Sci Rep (2013)

DosD positively regulates aerobic biofilm production in S. putrefaciens CN32.(A) Schematic of the genetic arrangement of DosD and its domain architecture. (B) Aerobic biofilm mass profiles of WT (white columns) and ΔdosD (gray columns) with simultaneous determination of growth of WT (solid line) and ΔdosD (dashed line). (C) Biofilm formation by ΔdosD complemented by vector-bearing dosD driven by its native promoter. The empty vector was transformed into WT and ΔdosD mutants as a control. (D) Attachment abilities of WT and ΔdosD. The average number of cells attached to the cover slips was counted in randomly picked areas of ten microscopic images. Each experiment was repeated at least three times and representative results are provided. Error bars refer to the standard deviation of the three samples.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3672883&req=5

f1: DosD positively regulates aerobic biofilm production in S. putrefaciens CN32.(A) Schematic of the genetic arrangement of DosD and its domain architecture. (B) Aerobic biofilm mass profiles of WT (white columns) and ΔdosD (gray columns) with simultaneous determination of growth of WT (solid line) and ΔdosD (dashed line). (C) Biofilm formation by ΔdosD complemented by vector-bearing dosD driven by its native promoter. The empty vector was transformed into WT and ΔdosD mutants as a control. (D) Attachment abilities of WT and ΔdosD. The average number of cells attached to the cover slips was counted in randomly picked areas of ten microscopic images. Each experiment was repeated at least three times and representative results are provided. Error bars refer to the standard deviation of the three samples.
Mentions: Genome annotation predicts that Sputcn32_3244 in S. putrefaciens CN32 encodes a putative diguanylate cyclase (DGC), composed of an N-terminal globin domain and a C-terminal GGDEF domain (Figure 1A). Given the typical function of the globin domain as a gas sensor16, we therefore postulated that Sputcn32_3244 might encode an oxygen sensing DGC (dosD) in S. putrefaciens CN32.

Bottom Line: DosD regulates the transcription of bpfA operon which encodes seven proteins including a large repetitive adhesin BpfA and its cognate type I secretion system (TISS).Regulation of DosD in aerobic biofilms is heavily dependent on an adhesin BpfA and the TISS.This study offers an insight into the molecular mechanism of oxygen-stimulated biofilm formation by S. putrefaciens CN32.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, University of Science & Technology of China, Hefei, China.

ABSTRACT
Although oxygen has been reported to regulate biofilm formation by several Shewanella species, the exact regulatory mechanism mostly remains unclear. Here, we identify a direct oxygen-sensing diguanylate cyclase (DosD) and reveal its regulatory role in biofilm formation by Shewanella putrefaciens CN32 under aerobic conditions. In vitro and in vivo analyses revealed that the activity of DosD culminates to synthesis of cyclic diguanylate (c-di-GMP) in the presence of oxygen. DosD regulates the transcription of bpfA operon which encodes seven proteins including a large repetitive adhesin BpfA and its cognate type I secretion system (TISS). Regulation of DosD in aerobic biofilms is heavily dependent on an adhesin BpfA and the TISS. This study offers an insight into the molecular mechanism of oxygen-stimulated biofilm formation by S. putrefaciens CN32.

Show MeSH
Related in: MedlinePlus