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The Global Response Regulator ExpA Controls Virulence Gene Expression through RsmA-Mediated and RsmA-Independent Pathways in Pectobacterium wasabiae SCC3193

机译:全局响应调节剂ExpA通过芥末油菜SCC3193中的RsmA介导和RsmA独立途径控制毒力基因表达。

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摘要

ExpA (GacA) is a global response regulator that controls the expression of major virulence genes, such as those encoding plant cell wall-degrading enzymes (PCWDEs) in the model soft rot phytopathogen Pectobacterium wasabiae SCC3193. Several studies with pectobacteria as well as related phytopathogenic gammaproteobacteria, such as Dickeya and Pseudomonas, suggest that the control of virulence by ExpA and its homologues is executed partly by modulating the activity of RsmA, an RNA-binding posttranscriptional regulator. To elucidate the extent of the overlap between the ExpA and RsmA regulons in P. wasabiae, we characterized both regulons by microarray analysis. To do this, we compared the transcriptomes of the wild-type strain, an expA mutant, an rsmA mutant, and an expA rsmA double mutant. The microarray data for selected virulence-related genes were confirmed through quantitative reverse transcription (qRT-PCR). Subsequently, assays were performed to link the observed transcriptome differences to changes in bacterial phenotypes such as growth, motility, PCWDE production, and virulence in planta. An extensive overlap between the ExpA and RsmA regulons was observed, suggesting that a substantial portion of ExpA regulation appears to be mediated through RsmA. However, a number of genes involved in the electron transport chain and oligogalacturonide metabolism, among other processes, were identified as being regulated by ExpA independently of RsmA. These results suggest that ExpA may only partially impact fitness and virulence via RsmA.
机译:ExpA(GacA)是一种全局响应调节剂,可控制主要毒力基因的表达,例如在模型软腐烂植物病原芥芥SCC3193中编码植物细胞壁降解酶(PCWDE)的那些基因。几项有关细菌和相关致病性γ-变形杆菌的研究,例如迪卡氏菌和假单胞菌,表明通过ExpA及其同系物控制毒力部分是通过调节RNA结合后转录调节因子RsmA的活性来实现的。为了阐明芥末假单胞菌ExpA和RsmA调控因子之间的重叠程度,我们通过微阵列分析对两种调控因子进行了表征。为此,我们比较了野生型菌株,expA突变体,rsmA突变体和expA rsmA双突变体的转录组。通过定量逆转录(qRT-PCR)确认了所选毒力相关基因的微阵列数据。随后,进行测定以将观察到的转录组差异与细菌表型的变化联系起来,例如生长,运动,PCWDE产生和植物中的毒力。观察到ExpA和RsmA调节子之间存在广泛的重叠,这表明ExpA调节的很大一部分似乎是通过RsmA介导的。但是,在其他过程中,许多与电子传输链和寡半乳糖醛酸代谢有关的基因被确定为受ExpA的独立于RsmA的调控。这些结果表明,ExpA可能仅部分影响RsmA的适应性和毒力。

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