RpoB 蛋白中的额外 L451G452N453 可抑制大肠杆菌在 37 度下因 DnaK/J 和触发因子耗竭而导致的合成致死率

IF 3.5 4区 生物学 Q2 MICROBIOLOGY Journal of Basic Microbiology Pub Date : 2024-09-17 DOI:10.1002/jobm.202400253
Dongjie Fan, Lushan Liu, Bella Yuen, Lu Sun, Yuliang Fu, Yan Liu, Rui Liao, Yanli Qu, Chuanpeng Liu, Qiming Zhou
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引用次数: 0

摘要

去除了伴侣触发因子和 DnaK/J 的大肠杆菌在 37°C 时无法存活,但在 30°C 以下时可以存活。在缺失触发因子和 DnaK/J 的工程大肠杆菌中,一种菌株 Z625 在 37 摄氏度下存活,而另一种菌株 Z629 仅在 30 摄氏度以下存活。对 Z625 和 Z629 的脂肪酸谱进行比较分析后发现,与野生型大肠杆菌 BW25113 相比,Z625 中缺乏大量饱和脂肪酸。此外,Z625 中存在更多的不饱和脂肪酸,而 Z629 的脂肪酸谱与 BW25113 非常相似。全基因组测序发现 Z625 的 rpoB 中有一个 9-bp 的插入物。对带有插入氨基酸序列 L451G452N453 的模拟 RpoB 蛋白的结构分析和对利福平的敏感性分析表明,插入的 RpoB 蛋白并没有扰乱其作为 RNA 聚合酶 beta 亚基的结构。对 Z625 和 Z629 进行的转录组学比较分析表明,这种插入影响了 Z625 中整个 RNA 聚合酶的转录,导致一些基因可能受到抑制,而这些基因的过度表达对大肠杆菌是有毒的。此外,Z625 表现出独特的新陈代谢适应性,这可能是其在 37°C 下存活的原因。总之,我们的研究阐明了 rpoB 中影响大肠杆菌转录调控的一个 LGN 插入,从而解释了缺失触发因子和 DnaK/J 的大肠杆菌在 37°C 下的存活。
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An Additional L451G452N453 in the RpoB Protein Suppressed the Synthetic Lethality in Escherichia coli at 37 Degrees Caused by Depletion of DnaK/J and Trigger Factor

Escherichia coli depletion of chaperone trigger factor and DnaK/J were not viable at 37°C, but viable below 30°C. Among the engineered E. coli depleted of trigger factor and DnaK/J, one strain Z625, exhibited survival at 37°C, while another strain Z629 only survived below 30°C. Comparative analysis of fatty acid profiles of Z625 and Z629 revealed absence of numerous saturated fatty acids in Z625 as compared to the wild-type E. coli BW25113. In addition, increased unsaturated fatty acids were present in Z625, whereas the fatty acids profile of Z629 closely resembled that of BW25113. Whole genome sequencing revealed a 9-bp insertion in rpoB of Z625. Combined structural analysis of simulated RpoB protein bearing the amino acid sequence L451G452N453 insertion and susceptibility analysis to rifampicin suggested that the insertion did not disturb the individual RpoB structure as beta subunit of RNA polymerase. Comparative transcriptomic analysis of Z625 and Z629 suggested that this insertion impacted transcription of the overall RNA polymerase in Z625, leading to potential repression of some genes whose overexpression was toxic to E. coli. Additionally, Z625 exhibited distinctive metabolic adaptations, likely contributing to its survival at 37°C. In summary, our study elucidated one LGN insertion in rpoB that impacts transcriptional regulation in E. coli, thereby explaining the survival of E. coli depletion of trigger factor and DnaK/J at 37°C, and these founding suggested that some simple mutations in critical genes like rpoB might play an important role in driving adaptive evolution.

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来源期刊
Journal of Basic Microbiology
Journal of Basic Microbiology 生物-微生物学
CiteScore
6.10
自引率
0.00%
发文量
134
审稿时长
1.8 months
期刊介绍: The Journal of Basic Microbiology (JBM) publishes primary research papers on both procaryotic and eucaryotic microorganisms, including bacteria, archaea, fungi, algae, protozoans, phages, viruses, viroids and prions. Papers published deal with: microbial interactions (pathogenic, mutualistic, environmental), ecology, physiology, genetics and cell biology/development, new methodologies, i.e., new imaging technologies (e.g. video-fluorescence microscopy, modern TEM applications) novel molecular biology methods (e.g. PCR-based gene targeting or cassettes for cloning of GFP constructs).
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