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Xanthomonas campestris sensor kinase HpaS co‐opts the orphan response regulator VemR to form a branched two‐component system that regulates motility

机译:Xanthomonas campestris传感器激酶HpaS与孤儿反应调节因子VemR共同形成分支的两组分系统调节运动

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摘要

pv. (Xcc) controls virulence and plant infection mechanisms via the activity of the sensor kinase and response regulator pair HpaS/hypersensitive response and pathogenicity G (HrpG). Detailed analysis of the regulatory role of HpaS has suggested the occurrence of further regulators besides HrpG. Here we used in vitro and in vivo approaches to identify the orphan response regulator VemR as another partner of HpaS and to characterize relevant interactions between components of this signalling system. Bacterial two‐hybrid and protein pull‐down assays revealed that HpaS physically interacts with VemR. Phos‐tag SDS‐PAGE analysis showed that mutation in reduced markedly the phosphorylation of VemR in vivo. Mutation analysis reveals that HpaS and VemR contribute to the regulation of motility and this relationship appears to be epistatic. Additionally, we show that VemR control of Xcc motility is due in part to its ability to interact and bind to the flagellum rotor protein FliM. Taken together, the findings describe the unrecognized regulatory role of sensor kinase HpaS and orphan response regulator VemR in the control of motility in Xcc and contribute to the understanding of the complex regulatory mechanisms used by Xcc during plant infection.
机译:pv。 (Xcc)通过传感器激酶和应答调节因子对HpaS /过敏反应和致病性G(HrpG)的活性来控制毒力和植物感染机制。对HpaS调节作用的详细分析表明,除了HrpG以外,还存在其他调节剂。在这里,我们使用了体外和体内方法,将孤儿应答调节因子VemR鉴定为HpaS的另一个伴侣,并表征了该信号系统组件之间的相关相互作用。细菌双杂交和蛋白质下拉分析表明,HpaS与VemR发生物理相互作用。磷酸标记SDS-PAGE分析表明,体内突变显着降低了VemR的磷酸化。突变分析显示,HpaS和VemR有助于调节运动,这种关系似乎是上位的。另外,我们表明,VemR对Xcc运动的控制部分是由于其与鞭毛转子蛋白FliM相互作用和结合的能力。综上所述,这些发现描述了未知的传感器激酶HpaS和孤儿反应调节因子VemR在Xcc运动控制中的调节作用,有助于理解Xcc在植物感染过程中所使用的复杂调节机制。

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