Novel iron-regulated and Fur-regulated small regulatory RNAs in Aggregatibacter actinomycetemcomitans
Novel iron-regulated and Fur-regulated small regulatory RNAs in Aggregatibacter actinomycetemcomitans
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DOI：
10.1111/j.2041-1014.2012.00645.x
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发表时间：
2012-10-01
影响因子：
3.7
通讯作者：
Haase, E. M.
中科院分区：
文献类型：
--
作者：
Amarasinghe, J. J.;Connell, T. D.;Haase, E. M.
铁可以通过非编码小RNA （sRNA）调控生物膜的形成。为了确定铁调控的sRNAs是否参与牙周病原聚集菌放线菌的生物膜形成，从添加铁或螯合培养的细菌中分离出总RNA。转录分析表明，生物信息学鉴定的4个sRNA分子（JA01JA04）在限铁培养基中的表达比富铁培养基显著上调。编码每个sRNA启动子的DNA片段能够在铁摄取调节剂滴定试验中从抑制毛皮的报告融合中滴定大肠杆菌铁摄取调节剂（Fur）。在凝胶转移实验中，含有重组AaFur的细胞裂解物改变了srna特异性dna的流动性。这些sRNAs的潜在靶标包括参与生物膜形成的基因。与过表达JA03 sRNA的放线菌相似的a .在琼脂上保持了粗糙的表型，但不再粘附在未包被的聚苯乙烯或玻璃上，尽管生物膜决定基因的表达仅轻微下降。综上所述，这些sRNAs具有调节生物膜形成的能力，但其功能靶基因仍有待证实。
Iron can regulate biofilm formation via non-coding small RNA (sRNA). To determine if iron-regulated sRNAs are involved in biofilm formation by the periodontopathogen Aggregatibacter actinomycetemcomitans, total RNA was isolated from bacteria cultured with iron supplementation or chelation. Transcriptional analysis demonstrated that the expression of four sRNA molecules (JA01JA04) identified by bioinformatics was significantly upregulated in iron-limited medium compared with iron-rich medium. A DNA fragment encoding each sRNA promoter was able to titrate Escherichia coli ferric uptake regulator (Fur) from a Fur-repressible reporter fusion in an iron uptake regulator titration assay. Cell lysates containing recombinant AaFur shifted the mobility of sRNA-specific DNAs in a gel shift assay. Potential targets of these sRNAs, determined in silico, included genes involved in biofilm formation. The A.similar to actinomycetemcomitans overexpressing JA03 sRNA maintained a rough phenotype on agar, but no longer adhered to uncoated polystyrene or glass, although biofilm determinant gene expression was only modestly decreased. In summary, these sRNAs have the ability to modulate biofilm formation, but their functional target genes remain to be confirmed.