Characteristics of the iron uptake-related process of a pathogenic Vibrio splendidus strain associated with massive mortalities of the sea cucumber Apostichopus japonicus
Characteristics of the iron uptake-related process of a pathogenic Vibrio splendidus strain associated with massive mortalities of the sea cucumber Apostichopus japonicus
复制标题
与海参大量死亡相关的致病性灿烂弧菌菌株铁吸收相关过程的特征
DOI：
10.1016/j.jip.2018.05.001
复制
发表时间：
2018
影响因子：
3.4
通讯作者：
Weiwei Zhang
中科院分区：
文献类型：
--
作者：
Tongxiang Song;Huijie Liu;Tengteng Lv;Xuelin Zhao;Yina Shao;Qingxi Han;Chenghua Li;Weiwei Zhang
在低铁生物利用率的环境下，许多细菌通过铁载体介导的铁获取系统获取铁用于生长和存活。然而，到目前为止，很少有关于在铁限制条件下灿烂弧菌的生长，铁载体的产生和铁载体受体的研究报道。在本研究中，灿烂弧菌可以在添加160 μM 2，2 ′-联吡啶（DIP）的培养基中存活，但在48 h时74.5%的生长受到抑制，而V. splendidusVs增加35.9%。作为OD 600 ofV.当DIP浓度从40 μM增加到80 μM时，splendidusVs的铁载体产量下降，spendidusVs在24 h时从34.0%增加到43.4%，并进一步确定其为异羟肟酸铁载体。目的：探索锚定在V外膜上的铁载体受体。在spendidusVs中，提取来自在80 μM DIP下和不含DIP下生长的细胞的外膜蛋白，并通过SDS-PAGE和MALDI-TOF/TOF MS鉴定差异表达的蛋白。鉴定了5种蛋白，即需氧菌素铁载体受体IutA、肠杆菌素受体蛋白FepA、ATP合酶亚基A、ATP合酶亚基B和ATP合酶F0 F1亚基β。实时荧光定量PCR结果显示，80 µM DIP处理6 h后，fiutA、fepA、atpA、atpBandatpβ-F0 F1的mRNA水平分别上调271.5、15.1、1.1、2.5和67.9倍。此外，铁载体受体基因fiutA和fepA的启动子具有明显的铁吸收调节因子（Fur）结合位点。结合异羟肟酸铁载体及其相应的需氧肌动蛋白铁载体受体IutA的同时产生，这些结果表明，在V中可能存在异羟肟酸铁载体IutA介导的铁摄取途径。辉煌的，辉煌的
Under low iron bioavailability environment, many bacteria acquire iron for growth and survival through siderophore-mediated iron acquisition systems. However, until now, little research on the growth, siderophore production and siderophore receptors ofVibrio splendidusVs under iron limited conditions has been reported. In our present study,V. splendidusVs could survive in media supplemented with 160 µM 2,2′-dipyridyl (DIP), but 74.5% of the growth was suppressed at 48 h, while the siderophore production ofV. splendidusVs increased by 35.9%. As the OD600ofV. splendidusVs decreased when the concentration of DIP was increased from 40 to 80 µM, the siderophore production ofV. splendidusVs increased from 34.0% to 43.4% at 24 h, and it was further determined to be a hydroxamate siderophore. To explore the potential siderophore receptors anchored on the outer membrane ofV. splendidusVs, outer membrane proteins from cells grown with and without 80 µM DIP were extracted and the differentially expressed proteins were identified by SDS-PAGE and MALDI-TOF/TOF MS. Five proteins, aerobactin siderophore receptor IutA, enterobactin receptor protein FepA, ATP synthase subunit A, ATP synthase subunit B and the ATP synthase F0F1 subunit beta were identified. Real-time reverse transcriptase PCR showed that mRNA levels ofiutA,fepA, atpA,atpBandatpβ-F0F1were upregulated 271.5-, 15.1-, 1.1-, 2.5- and 67.9-fold respectively, after 6 h in cells treated with 80 µM DIP. In addition, the promoters of the siderophore receptor genes ofiutAandfepAhad apparent ferric uptake regulator (Fur) binding sites. Combined with the simultaneous production of both the hydroxamate siderophore and its corresponding aerobactin siderophore receptor IutA, these results suggested that there might be a hydroxamate siderophore-IutA mediated iron uptake pathway inV. splendidusVs.