Dominelli Nazzareno, Regaiolo Alice, Willy Leon, Heermann Ralf
Institute of Molecular Physiology (imP), Johannes-Gutenberg-University Mainz, Biocenter II, Microbiology and Biotechnology, Hanns-Dieter-Hüsch-Weg 17, 55128 Mainz, Germany.
Institute for Biotechnology and Drug Research gGmbH (IBWF), Hanns-Dieter-Hüsch-Weg 17, 55128 Mainz, Germany.
Microorganisms. 2023 Mar 30;11(4):890. doi: 10.3390/microorganisms11040890.
In bacteria, group-coordinated behavior such as biofilm formation or virulence are often mediated via cell-cell communication, a process referred to as quorum sensing (QS). The canonical QS system of Gram-negative bacteria uses -acyl homoserine lactones (AHLs) as communication molecules, which are produced by LuxI-type synthases and sensed by cognate LuxR-type receptors. These receptors act as transcriptional regulators controlling the expression of specific genes. Some bacteria harbor LuxR-type receptors lacking a cognate LuxI-type synthases, designated as LuxR solos. Among many other LuxR solos, the entomopathogenic enteric bacterium harbors a SdiA-like LuxR solo containing an AHL signal-binding domain, for which a respective signal molecule and target genes have not been identified yet. Here we performed SPR analysis to demonstrate that SdiA acts as a bidirectional regulator of transcription, tightly controlling its own expression and the adjacent () gene in a gene supposed to be involved in the colonization of eukaryotes. Via qPCR we could further determine that in deletion mutant strains, is upregulated, indicating that SdiA negatively affects expression of . Furthermore, the Δ deletion mutant exhibited differences in biofilm formation and motility compared with the wild-type. Finally, using nanoDSF analysis we could identify putative binding ability of SdiA towards diverse AHLs, but also to plant-derived signals, modulating the DNA-binding capacity of SdiA, suggesting that this LuxR solo acts as an important player in interkingdom signaling between and plants.
在细菌中,群体协调行为如生物膜形成或毒力通常通过细胞间通讯介导,这一过程称为群体感应(QS)。革兰氏阴性菌的经典QS系统使用N-酰基高丝氨酸内酯(AHLs)作为通讯分子,由LuxI型合成酶产生,并由同源的LuxR型受体感知。这些受体作为转录调节因子,控制特定基因的表达。一些细菌含有缺乏同源LuxI型合成酶的LuxR型受体,称为孤独型LuxR(LuxR solos)。在许多其他孤独型LuxR中,昆虫病原性肠道细菌含有一种类似SdiA的孤独型LuxR,其含有一个AHL信号结合结构域,尚未鉴定出相应的信号分子和靶基因。在这里,我们进行了表面等离子体共振(SPR)分析,以证明SdiA作为转录的双向调节因子,紧密控制其自身表达以及在一种被认为参与真核生物定殖的基因中的相邻基因的表达。通过定量聚合酶链反应(qPCR),我们可以进一步确定,在SdiA缺失突变株中,该基因上调,表明SdiA对该基因的表达有负面影响。此外,与野生型相比,ΔSdiA缺失突变体在生物膜形成和运动性方面表现出差异。最后,使用纳米差示扫描荧光法(nanoDSF)分析,我们可以确定SdiA对多种AHLs以及植物衍生信号的假定结合能力,调节SdiA的DNA结合能力,这表明这种孤独型LuxR在昆虫病原性肠道细菌与植物之间的跨界信号传导中起重要作用。