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bolA gene involved in curli amyloids and fimbriae production in E. coli: exploring pathways to inhibit biofilm and amyloid formation
Journal of Biological Research-Thessaloniki ( IF 3.3 ) Pub Date : 2020-06-17 , DOI: 10.1186/s40709-020-00120-7
Mohd W. Azam , Azna Zuberi , Asad U. Khan

Biofilm formation is a complex phenomenon of bacterial cells, involved in several human infections. Its formation is regulated and controlled by several protein factors. The BolA-like proteins (bolA gene) are conserved in both prokaryotes and eukaryotes. The BolA protein is a transcription factor involved in bacterial cell motility and biofilm formation. This study was initiated to elucidate the role of the bolA gene in the curli biogenesis and amyloid production as well as to observe changes in the expression of fimH, a fimbriae gene. Knockdown mutants of Escherichia coli MG1655 bolA gene (bolA-KD) were generated using CRISPR interference. The results obtained, were validated through gene expression using RT-PCR, microscopic analysis and different biofilm and amyloid assays. The bolA knockdown mutants showed a decrement in curli amyloid fibers, in fimbriae production and biofilm formation. We have also observed a reduction in EPS formation, eDNA production and extracellular protein content. Gene expression data showed that bolA downregulation caused the suppression of csgA and csgD of curli that led to the reduction in curli fiber and the amyloid formation and also the suppression of fimH, leading to the loss of fimbriae. Curli fibers and fimbriae are found to be involved in biofilm formation leading to the pathogenicity of the bacterial cell. BolA is a conserved protein and is found to play a significant role in curli and fimbriae formation in E. coli. This study further proved that CRISPRi mediated suppression of the bolA gene leads to inhibition of biofilm formation through curli and fimbriae inhibition. Hence, it may be proposed as a possible target for intervention of biofilm mediated infections.

中文翻译:

bolA基因参与大肠杆菌中的卷曲淀粉样蛋白和毛生成:探索抑制生物膜和淀粉样蛋白形成的途径

生物膜的形成是细菌细胞的复杂现象,涉及多种人类感染。它的形成受几种蛋白质因子的调控。BolA样蛋白(bolA基因)在原核生物和真核生物中均保守。BolA蛋白是参与细菌细胞运动和生物膜形成的转录因子。开展这项研究是为了阐明bolA基因在卷曲生物的发生和淀粉样蛋白产生中的作用,以及观察fimH(一种菌毛基因)表达的变化。大肠杆菌MG1655 bolA基因的敲低突变体(bolA-KD)使用CRISPR干扰产生。获得的结果通过使用RT-PCR的基因表达,显微镜分析以及不同的生物膜和淀粉样蛋白测定法得到验证。bolA敲低突变体显示出卷曲的淀粉样蛋白纤维减少,在菌丝生产和生物膜形成中。我们还观察到EPS形成,eDNA产生和细胞外蛋白质含量的减少。基因表达数据表明,bolA的下调导致对curli的csgA和csgD的抑制,从而导致curli纤维的减少和淀粉样蛋白的形成以及fimH的抑制,从而导致菌毛的丢失。发现卷毛纤维和菌毛参与生物膜的形成,导致细菌细胞的致病性。BolA是一种保守的蛋白质,在大肠杆菌中的卷曲和菌毛形成中起着重要作用。这项研究进一步证明,CRISPRi介导的bolA基因抑制可通过卷曲和菌毛抑制作用抑制生物膜形成。因此,
更新日期:2020-07-24
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