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首页> 外文期刊>Journal of bacteriology >The Cyclic-di-GMP Phosphodiesterase BinA Negatively Regulates Cellulose-Containing Biofilms in Vibrio fischeri
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The Cyclic-di-GMP Phosphodiesterase BinA Negatively Regulates Cellulose-Containing Biofilms in Vibrio fischeri

机译:环二GMP磷酸二酯酶BinA负调节费氏弧菌中的纤维素生物膜。

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Bacteria produce different types of biofilms under distinct environmental conditions. Vibrio fischeri has the capacity to produce at least two distinct types of biofilms, one that relies on the symbiosis polysaccharide Syp and another that depends upon cellulose. A key regulator of biofilm formation in bacteria is the intracellular signaling molecule cyclic diguanylate (c-di-GMP). In this study, we focused on a predicted c-di-GMP phosphodiesterase encoded by the gene binA, located directly downstream of syp, a cluster of 18 genes critical for biofilm formation and the initiation of symbiotic colonization of the squid Euprymna scolopes. Disruption or deletion of binA increased biofilm formation in culture and led to increased binding of Congo red and calcofluor, which are indicators of cellulose production. Using random transposon mutagenesis, we determined that the phenotypes of the ΔbinA mutant strain could be disrupted by insertions in genes in the bacterial cellulose biosynthesis cluster (bcs), suggesting that cellulose production is negatively regulated by BinA. Replacement of critical amino acids within the conserved EAL residues of the EAL domain disrupted BinA activity, and deletion of binA increased c-di-GMP levels in the cell. Together, these data support the hypotheses that BinA functions as a phosphodiesterase and that c-di-GMP activates cellulose biosynthesis. Finally, overexpression of the syp regulator sypG induced binA expression. Thus, this work reveals a mechanism by which V. fischeri inhibits cellulose-dependent biofilm formation and suggests that the production of two different polysaccharides may be coordinated through the action of the cellulose inhibitor BinA.
机译:细菌在不同的环境条件下会产生不同类型的生物膜。 费氏弧菌具有生产至少两种不同类型生物膜的能力,一种依赖共生多糖Syp,另一种依赖纤维素。细菌中生物膜形成的关键调控因子是细胞内信号分子环状双鸟苷酸(c-di-GMP)。在这项研究中,我们重点研究了由 binA 基因编码的预测的c-di-GMP磷酸二酯酶,该基因直接位于 syp 的下游,该基因是对生物膜形成至关重要的18个基因的簇鱿鱼 Euprymna scolopes 的共生定居和启动。 binA 的破坏或缺失增加了培养物中生物膜的形成,并导致刚果红和calcofluor的结合增加,这是纤维素生产的指标。使用随机转座子诱变,我们确定Δ binA 突变株的表型可以通过插入细菌纤维素生物合成簇( bcs )中的基因来破坏,这表明纤维素生产受到BinA的负面监管。 EAL域的保守EAL残基内关键氨基酸的置换破坏BinA活性,而 binA 的缺失增加了细胞中c-di-GMP的水平。总之,这些数据支持以下假设:BinA充当磷酸二酯酶,而c-di-GMP激活纤维素生物合成。最后, syp 调节剂 sypG 的过表达诱导 binA 表达。因此,这项工作揭示了 V的机制。 fischeri 抑制了纤维素依赖性生物膜的形成,并表明可能通过纤维素抑制剂BinA的作用来协调两种不同多糖的产生。

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