Specificity of Subtilin-Mediated Activation of Histidine Kinase SpaK
Specificity of Subtilin-Mediated Activation of Histidine Kinase SpaK
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DOI：
10.1128/aem.00781-17
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发表时间：
2017-07
影响因子：
4.4
通讯作者：
Christoph Geiger;T. Spieß;S. Korn;P. Kötter;K. Entian
中科院分区：
文献类型：
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作者：
Christoph Geiger;T. Spieß;S. Korn;P. Kötter;K. Entian
摘要通过双组分系统的自诱导是一种广泛存在的感知环境和代谢变化的调节机制。虽然羊毛硫抗生素乳链菌肽和枯草菌素密切相关，并共享相同的羊毛硫环结构，但它们以高度特异性的方式自诱导其生物合成。枯草菌素仅激活枯草芽孢杆菌的双组分系统SpaRK，而乳链菌肽仅激活乳酸乳球菌的双组分系统NisRK。为了鉴定决定枯草杆菌素自身诱导特异性的组分，分析了相应羊毛硫抗生素的几种变体的自身诱导能力。在这里，我们表明，氨基酸位置20是至关重要的SpaK激活，作为一个工程化的乳酸链球菌素分子与苯丙氨酸在位置20（乳酸链球菌素N20 F）能够激活SpaK在一个特定的方式。与枯草杆菌素N-末端色氨酸（nisin I1 W/N20 F）组合，SpaK的自诱导几乎达到枯草杆菌素介导的自诱导水平。此外，枯草杆菌蛋白的整体结构对于其与组氨酸激酶的关联也是重要的。破坏第二个lanthirons环（枯草杆菌素C11 A，环B），以及突变，干扰位于lanthirons环C和D之间的铰链区的灵活性（枯草杆菌素L21 P/Q22 P），废除SpaK自身诱导。虽然枯草杆菌素的C-末端部分是有效的SpaK自诱导所必需的，但对羊毛硫醚环D和E的破坏没有可测量的影响。基于这些发现，枯草杆菌素与组氨酸激酶SpaK的相互作用的模型建立。重要性虽然双组分系统是重要的调节系统，传感环境变化，非常少的信息传感器的分子机制或相互作用的传感器与其各自的激酶。线性羊毛硫抗生素如枯草菌素和乳链菌肽对它们各自的激酶的强特异性提供了极好的模型系统来阐明这些羊毛硫抗生素以特定方式激活组氨酸激酶的结构需求。不仅如此，羊毛硫抗生素的生物合成是通过双组分系统自诱导的。因此，了解它们与组氨酸激酶的相互作用是新工程肽抗生素的生物合成所必需的。使用基于枯草芽孢杆菌的报告系统，我们能够确定特定SpaK激活所必需的分子限制，并仅用两个点突变为乳链菌肽提供SpaK特异性。
ABSTRACT Autoinduction via two-component systems is a widespread regulatory mechanism that senses environmental and metabolic changes. Although the lantibiotics nisin and subtilin are closely related and share the same lanthionine ring structure, they autoinduce their biosynthesis in a highly specific manner. Subtilin activates only the two-component system SpaRK of Bacillus subtilis, whereas nisin activates solely the two-component system NisRK of Lactococcus lactis. To identify components that determine the specificity of subtilin autoinduction, several variants of the respective lantibiotics were analyzed for their autoinductive capacities. Here, we show that amino acid position 20 is crucial for SpaK activation, as an engineered nisin molecule with phenylalanine at position 20 (nisin N20F) was able to activate SpaK in a specific manner. In combination with the N-terminal tryptophan of subtilin (nisin I1W/N20F), SpaK autoinduction reached almost the level of subtilin-mediated autoinduction. Furthermore, the overall structure of subtilin is also important for its association with the histidine kinase. The destruction of the second lanthionine ring (subtilin C11A, ring B), as well as mutations that interfere with the flexibility of the hinge region located between lanthionine rings C and D (subtilin L21P/Q22P), abolished SpaK autoinduction. Although the C-terminal part of subtilin is needed for efficient SpaK autoinduction, the destruction of lanthionine rings D and E had no measurable impact. Based on these findings, a model for the interaction of subtilin with histidine kinase SpaK was established. IMPORTANCE Although two-component systems are important regulatory systems that sense environmental changes, very little information on the molecular mechanism of sensing or the interaction of the sensor with its respective kinase is available. The strong specificity of linear lantibiotics such as subtilin and nisin for their respective kinases provides an excellent model system to unravel the structural needs of these lantibiotics for activating histidine kinases in a specific manner. More than that, the biosyntheses of lantibiotics are autoinduced via two-component systems. Therefore, an understanding of their interactions with histidine kinases is needed for the biosynthesis of newly engineered peptide antibiotics. Using a Bacillus subtilis-based reporter system, we were able to identify the molecular constraints that are necessary for specific SpaK activation and to provide SpaK specificity to nisin with just two point mutations.