Transcript Profiling of Nitroxoline-Treated Biofilms Shows Rapid Up-regulation of Iron Acquisition Gene Clusters.
Transcript Profiling of Nitroxoline-Treated Biofilms Shows Rapid Up-regulation of Iron Acquisition Gene Clusters.
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DOI：
10.1021/acsinfecdis.2c00206
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发表时间：
2022-08-12
影响因子：
5.3
通讯作者：
Huigens, Robert W., III
中科院分区：
文献类型：
--
作者：
Liu, Ke;Abouelhassan, Yasmeen;Zhang, Yanping;Jin, Shouguang;Huigens, Robert W., III
细菌生物膜是一种由生物分子组成的保护性基质，包埋在细菌表面的缓慢或非复制细胞群落。与自由漂浮的浮游细菌不同，生物膜天生对常规抗生素具有耐受性，并且在复发性和慢性感染中普遍存在。硝基喹啉，一种广谱的生物膜根除剂，被用来探测生物膜的活力。转录谱分析（RNA-seq）显示，耐甲氧西林金黄色葡萄球菌（MRSA）生物膜中的2594个基因中有452个（17.4%）在硝羟喹啉处理2 h后差异表达。WoPPER分析和时间过程验证（RT-qPCR）揭示，参与铁获取的基因簇（sbn、isd、MW 2101、MW 0695、fhu和feo）在硝羟喹啉处理后快速上调，这指示MRSA生物膜中的铁饥饿。此外，寡肽转运蛋白和核黄素生物合成相关基因表达上调，而类胡萝卜素生物合成和硝酸盐同化相关基因表达下调。RT-qPCR实验表明，铁摄取转录也上调建立表皮葡萄球菌和鲍曼不动杆菌生物膜硝羟喹啉治疗后。总的来说，我们表明RNA-seq是一个理想的平台，可以定义对生物膜存活至关重要的细胞途径，此外还证明了这些细菌群落对铁的需求。
Bacterial biofilms are surface-attached communities of slow- or non-replicating cells embedded within a protective matrix of biomolecules. Unlike free-floating planktonic bacteria, biofilms are innately tolerant to conventional antibiotics and are prevalent in recurring and chronic infections. Nitroxoline, a broad-spectrum biofilm-eradicating agent, was used to probe biofilm viability. Transcript profiling (RNA-seq) showed that 452 of 2594 genes (17.4%) in methicillin-resistant Staphylococcus aureus (MRSA) biofilms were differentially expressed after a 2 h treatment of nitroxoline. WoPPER analysis and time-course validation (RT-qPCR) revealed that gene clusters involved in iron acquisition (sbn, isd, MW2101, MW0695, fhu, and feo) were rapidly up-regulated following nitroxoline treatment, which is indicative of iron starvation in MRSA biofilms. In addition, genes related to oligopeptide transporters and riboflavin biosynthesis were found to be up-regulated, while genes related to carotenoid biosynthesis and nitrate assimilation were down-regulated. RT-qPCR experiments revealed that iron uptake transcripts were also up-regulated in established Staphylococcus epidermidis and Acinetobacter baumannii biofilms following nitroxoline treatment. Overall, we show RNA-seq to be an ideal platform to define cellular pathways critical for biofilm survival, in addition to demonstrating the need these bacterial communities have for iron.
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