1887

Abstract

In enterobacteria, the CsgD protein activates production of two extracellular structures: thin aggregative fimbriae (curli) and cellulose. While curli fibres promote biofilm formation and cell aggregation, the evidence for a direct role of cellulose as an additional determinant for biofilm formation is not as straightforward. The MG1655 laboratory strain of only produces limited amounts of curli and cellulose; however, ectopic expression results in strong stimulation of curli and cellulose production. We show that, in a -overexpressing derivative of MG1655, cellulose production negatively affects curli-mediated surface adhesion and cell aggregation, thus acting as a negative determinant for biofilm formation. Consistent with this observation, deletion of the gene, necessary for cellulose production, resulted in a significant increase in curli-dependent adhesion. We found that cellulose production increased tolerance to desiccation, suggesting that the function of cellulose might be related to resistance to environmental stresses rather than to biofilm formation. Production of the curli/cellulose network in enterobacteria typically takes place at low growth temperature (<32 °C), but not at 37 °C. We show that CsgD overexpression can overcome temperature-dependent control of the curli-encoding operon, but not of the cellulose-related gene, suggesting very tight temperature control of cellulose production in MG1655.

Keyword(s): CV, crystal violet
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2008-07-01
2019-11-21
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Phenotypes of strains MG1655 (WT), PHL856 ( ) and LG26 (Δ ) on Congo Red-supplemented agar medium (CR medium), either at 30°C (upper panel) or at 37°C (lower panel). Strains were transformed either with the pT7-7 control vector or with the pT7CsgD plasmid as indicated. [ PDF] (29 kb) SDS-PAGE of formic-acid-treated membrane fractions of MG1655 (WT); PHL856 ( :: ); LG26 (Δ :: ) transformed either with the pT7-7 or with the pT7CsgD plasmid, grown at 30°C. The band indicated by the arrow has an apparent molecular mass of 17 kDa, consistent with the molecular mass of CsgA, the curli major subunit. [ PDF] (36 kb)

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Phenotypes of strains MG1655 (WT), PHL856 ( ) and LG26 (Δ ) on Congo Red-supplemented agar medium (CR medium), either at 30°C (upper panel) or at 37°C (lower panel). Strains were transformed either with the pT7-7 control vector or with the pT7CsgD plasmid as indicated. [ PDF] (29 kb) SDS-PAGE of formic-acid-treated membrane fractions of MG1655 (WT); PHL856 ( :: ); LG26 (Δ :: ) transformed either with the pT7-7 or with the pT7CsgD plasmid, grown at 30°C. The band indicated by the arrow has an apparent molecular mass of 17 kDa, consistent with the molecular mass of CsgA, the curli major subunit. [ PDF] (36 kb)

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