Department of Microbiology, University of Washington, Seattle, WA 98195.
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
Proc Natl Acad Sci U S A. 2018 Sep 25;115(39):9785-9790. doi: 10.1073/pnas.1809611115. Epub 2018 Sep 6.
Certain plant-associated Proteobacteria sense their host environment by detecting an unknown plant signal recognized by a member of a LuxR subfamily of transcription factors. This interkingdom communication is important for both mutualistic and pathogenic interactions. The root endophyte sp. GM79 possesses such a regulator, named PipR. In a previous study we reported that PipR activates an adjacent gene () coding for a proline iminopeptidase in response to leaf macerates and peptides and that this activation is dependent on a putative ABC-type transporter [Schaefer AL, et al. (2016) mBio 7:e01101-16]. In this study we identify a chemical derived from ethanolamine that induces PipR activity at picomolar concentrations, and we present evidence that this is the active inducer present in plant leaf macerates. First, a screen of more than 750 compounds indicated ethanolamine was a potent inducer for the PipR-sensing system; however, ethanolamine failed to bind to the periplasmic-binding protein (PBP) required for the signal response. This led us to discover that a specific ethanolamine derivative, -(2-hydroxyethyl)-2-(2-hydroxyethylamino) acetamide (HEHEAA), binds to the PBP and serves as a potent PipR-dependent inducer. We also show that a compound, which coelutes with HEHEAA in HPLC and induces gene expression in a PipR-dependent manner, can be found in leaf macerates. This work sheds light on how plant-associated bacteria can sense their environment and on the nature of inducers for a family of plant-responsive LuxR-like transcription factors found in plant-associated bacteria.
某些与植物相关的变形菌通过检测被转录因子 LuxR 亚家族成员识别的未知植物信号来感知其宿主环境。这种种间通讯对于互利共生和致病相互作用都很重要。根内生菌 sp. GM79 拥有这样的调节剂,称为 PipR。在之前的研究中,我们报道 PipR 会响应叶提取物和肽激活相邻基因 (),该基因编码脯氨酸亚氨基肽酶,并且这种激活依赖于假定的 ABC 型转运体 [Schaefer AL, 等人。(2016)mBio 7:e01101-16]。在这项研究中,我们鉴定出一种从乙醇胺衍生而来的化学物质,可在皮摩尔浓度下诱导 PipR 活性,并提供证据表明该物质是植物叶提取物中存在的有效诱导剂。首先,对超过 750 种化合物的筛选表明乙醇胺是 PipR 感应系统的有效诱导剂;然而,乙醇胺未能与信号响应所需的周质结合蛋白 (PBP) 结合。这导致我们发现一种特定的乙醇胺衍生物,-(2-羟乙基)-2-(2-羟乙基氨基)乙酰胺 (HEHEAA),与 PBP 结合并作为一种有效的 PipR 依赖性诱导剂。我们还表明,在 HPLC 中与 HEHEAA 共洗脱并以 PipR 依赖性方式诱导 基因表达的化合物可以在 叶提取物中找到。这项工作揭示了与植物相关的细菌如何感知其环境,以及植物相关细菌中发现的一类植物响应 LuxR 样转录因子的诱导剂的性质。