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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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The bpfA operon is required for DosD-induced biofilm enhancement.Biofilms formed by ΔbpfA, ΔdosDbpfA, ΔaggA and ΔdosDaggA after 12 h of growth. Representative glass tubes after stained by crystal violet for biofilm assay are shown on the top.
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f5: The bpfA operon is required for DosD-induced biofilm enhancement.Biofilms formed by ΔbpfA, ΔdosDbpfA, ΔaggA and ΔdosDaggA after 12 h of growth. Representative glass tubes after stained by crystal violet for biofilm assay are shown on the top.

Mentions: The role of BpfA and AggA in DosD-regulated aerobic biofilm formation was subsequently evaluated. In this case, bpfA and aggA were deleted in the WT and the ΔdosD, respectively. BpfA in S. putrefaciens CN32 is a large protein with a molecular weight of 429 KDa with repetitive sequences. To thoroughly deactivate bpfA, primers were designed to introduce a frameshift mutation which produced ~250 stop codons in the remaining sequence of the bpfA in genome. The biofilm analysis shows that ΔbpfA, ΔdosDΔbpfA, ΔaggA and ΔdosDΔaggA had similar and greatly impaired biofilm formation compared with WT (Figure 5). This result suggests that BpfA is the primary contributor to DosD-regulated biofilm formation. AggA might not directly contribute to DosD-regulated biofilm formation, but rather act indirectly by affecting BpfA secretion.


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)

The bpfA operon is required for DosD-induced biofilm enhancement.Biofilms formed by ΔbpfA, ΔdosDbpfA, ΔaggA and ΔdosDaggA after 12 h of growth. Representative glass tubes after stained by crystal violet for biofilm assay are shown on the top.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: The bpfA operon is required for DosD-induced biofilm enhancement.Biofilms formed by ΔbpfA, ΔdosDbpfA, ΔaggA and ΔdosDaggA after 12 h of growth. Representative glass tubes after stained by crystal violet for biofilm assay are shown on the top.
Mentions: The role of BpfA and AggA in DosD-regulated aerobic biofilm formation was subsequently evaluated. In this case, bpfA and aggA were deleted in the WT and the ΔdosD, respectively. BpfA in S. putrefaciens CN32 is a large protein with a molecular weight of 429 KDa with repetitive sequences. To thoroughly deactivate bpfA, primers were designed to introduce a frameshift mutation which produced ~250 stop codons in the remaining sequence of the bpfA in genome. The biofilm analysis shows that ΔbpfA, ΔdosDΔbpfA, ΔaggA and ΔdosDΔaggA had similar and greatly impaired biofilm formation compared with WT (Figure 5). This result suggests that BpfA is the primary contributor to DosD-regulated biofilm formation. AggA might not directly contribute to DosD-regulated biofilm formation, but rather act indirectly by affecting BpfA secretion.

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