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首页> 外文期刊>Journal of bacteriology >Isolation and Characterization of a Shewanella putrefaciens MR-1 Electron Transport RegulatoretrA Mutant: Reassessment of the Role of EtrA
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Isolation and Characterization of a Shewanella putrefaciens MR-1 Electron Transport RegulatoretrA Mutant: Reassessment of the Role of EtrA

机译：腐烂希瓦氏菌MR-1电子传输调节剂A突变体的分离和表征：EtrA的作用的重新评估。

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Shewanella putrefaciens MR-1 has emerged as a good model to study anaerobic respiration and electron transport-linked metal reduction. Its remarkable respiratory plasticity suggests the potential for a complex regulatory system to coordinate electron acceptor use in the absence of O2. It had previously been suggested that EtrA (electron transport regulator A), an analog of Fnr (fumarate nitrate regulator) from Escherichia coli, may regulate gene expression for anaerobic electron transport. AnetrA knockout strain (ETRA-153) was isolated from MR-1 using a gene replacement strategy. Reverse transcription-PCR analysis of total RNA demonstrated the loss of the etrAmRNA in ETRA-153. ETRA-153 cells retained the ability to grow on all electron acceptors tested, including fumarate, trimethylamineN-oxide (TMAO), thiosulfate, dimethyl sulfoxide, ferric citrate, nitrate, and O2, as well as the ability to reduce ferric citrate, manganese(IV), nitrate, and nitrite. EtrA is therefore not necessary for growth on, or the reduction of, these electron acceptors. However, ETRA-153 had reduced initial growth rates on fumarate and nitrate but not on TMAO. The activities for fumarate and nitrate reductase were lower in ETRA-153, as were the levels of fumarate reductase protein and transcript. ETRA-153 was also deficient in one type of ubiquinone. These results are in contrast to those previously reported for the putative etrA mutant METR-1. Molecular analysis of METR-1 indicated that its etrAgene is not interrupted; its reported phenotype was likely due to the use of inappropriate anaerobic growth conditions.
机译：腐臭希瓦氏菌 MR-1已成为研究厌氧呼吸和电子传输相关金属还原的良好模型。其出色的呼吸可塑性表明，在缺乏O 2 的情况下，可能需要复杂的调控系统来协调电子受体的使用。以前曾有人提出，EtrA（电子转运调节剂A）是大肠杆菌的Fnr（富马酸酯硝酸盐调节剂）的类似物，可以调节厌氧电子转运的基因表达。使用基因替换策略从MR-1中分离出一个 etrA 敲除菌株（ETRA-153）。总RNA的反转录PCR分析表明，ETRA-153中的 etrA mRNA丢失。 ETRA-153细胞保留了在所有测试的电子受体上生长的能力，这些电子受体包括富马酸盐，三甲胺 N -氧化物（TMAO），硫代硫酸盐，二甲基亚砜，柠檬酸铁，硝酸盐和O 2 < / sub>，以及还原柠檬酸铁，锰（IV），硝酸盐和亚硝酸盐的能力。因此，EtrA对于在这些电子受体上生长或还原不需要。但是，ETRA-153降低了富马酸酯和硝酸盐的初始生长速率，但未降低TMAO的初始生长速率。富马酸酯和硝酸还原酶的活性在ETRA-153中较低，富马酸酯还原酶蛋白和转录水平也较低。 ETRA-153也缺乏一种泛醌。这些结果与先前报道的假定的 etrA 突变体METR-1的结果相反。分子分析表明，METR-1的 etrA 基因没有被打断。其报告的表型可能是由于使用了不合适的厌氧生长条件。

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