Characterization of ExeM, an Extracellular Nuclease of Shewanella oneidensis MR-1.
Characterization of ExeM, an Extracellular Nuclease of Shewanella oneidensis MR-1.
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DOI：
10.3389/fmicb.2018.01761
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发表时间：
2018
影响因子：
5.2
通讯作者：
Thormann KM
中科院分区：
文献类型：
--
作者：
Binnenkade L;Kreienbaum M;Thormann KM
细菌胞外核酸酶在多种过程中具有多种功能，如营养获取、自然转化、生物膜形成或防御中性粒细胞胞外陷阱（NETs）。本文研究了舍瓦氏菌MR-1（一种胞外核酸酶）中ExeM的特性，该酶在舍瓦氏菌、弧菌、气单胞菌等物种中广泛保守。在s.o oneidensis中，ExeM对正常的生物膜形成至关重要。对异种产生的ExeM的体外活性测量表明，该酶是一种糖非特异性内切酶，需要Ca2+和Mg2+/Mn2+作为辅助因子才能充分发挥活性。即使当假定的c端膜锚点被删除时，ExeM几乎完全定位于细胞质膜部分。相反，在培养基上清液中未检测到ExeM。基于这些结果，我们假设ExeM主要与细胞附近的DNA相互作用，例如促进生物膜的形成和防御NETs，或控制DNA的摄取。
Bacterial extracellular nucleases have multiple functions in processes as diverse as nutrient acquisition, natural transformation, biofilm formation, or defense against neutrophil extracellular traps (NETs). Here we explored the properties of ExeM in Shewanella oneidensis MR-1, an extracellular nuclease, which is widely conserved among species of Shewanella, Vibrio, Aeromonas, and others. In S. oneidensis, ExeM is crucial for normal biofilm formation. In vitro activity measurements on heterologously produced ExeM revealed that this enzyme is a sugar-unspecific endonuclease, which requires Ca2+ and Mg2+/Mn2+ as co-factors for full activity. ExeM was almost exclusively localized to the cytoplasmic membrane fraction, even when a putative C-terminal membrane anchor was deleted. In contrast, ExeM was not detected in medium supernatants. Based on the results we hypothesize that ExeM predominantly interacts with DNA in close proximity to the cell, e.g., to promote biofilm formation and defense against NETs, or to control uptake of DNA.
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