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A cyclic GMP‐dependent signalling pathway regulates bacterial phytopathogenesis

机译:a cyclic Gmp‐dependent signalling pathway regulates bacterial phytopathogenesis

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摘要

Cyclic guanosine 3′,5′‐monophosphate (cyclic GMP) is a second messenger whose role in bacterial signalling is poorly understood. A genetic screen in the plant pathogen Xanthomonas campestris (Xcc) identified that XC_0250, which encodes a protein with a class III nucleotidyl cyclase domain, is required for cyclic GMP synthesis. Purified XC_0250 was active in cyclic GMP synthesis in vitro. The linked gene XC_0249 encodes a protein with a cyclic mononucleotide‐binding (cNMP) domain and a GGDEF diguanylate cyclase domain. The activity of XC_0249 in cyclic di‐GMP synthesis was enhanced by addition of cyclic GMP. The isolated cNMP domain of XC_0249 bound cyclic GMP and a structure–function analysis, directed by determination of the crystal structure of the holo‐complex, demonstrated the site of cyclic GMP binding that modulates cyclic di‐GMP synthesis. Mutation of either XC_0250 or XC_0249 led to a reduced virulence to plants and reduced biofilm formation in vitro. These findings describe a regulatory pathway in which cyclic GMP regulates virulence and biofilm formation through interaction with a novel effector that directly links cyclic GMP and cyclic di‐GMP signalling.
机译:环状鸟苷3',5'-单磷酸酯(环状GMP)是第二信使,人们对它在细菌信号传导中的作用了解甚少。植物病原体黄单胞菌(Xanthomonas campestris,Xcc)的遗传筛选确定,循环GMP合成需要XC_0250,它编码具有III类核苷酸环化酶结构域的蛋白质。纯化的XC_0250在体外可用于循环GMP合成。连锁基因XC_0249编码具有环状单核苷酸结合(cNMP)结构域和GGDEF二鸟苷酸环化酶结构域的蛋白质。 XC_0249在环状di-GMP合成中的活性通过添加环状GMP得以增强。 XC_0249的分离的cNMP结构域结合了环状GMP,并通过确定整体复合物的晶体结构进行了结构-功能分析,证明了环状GMP结合的位点调节了环di-GMP的合成。 XC_0250或XC_0249的突变导致对植物的毒力降低和体外生物膜形成减少。这些发现描述了一条调控途径,其中环状GMP通过与直接连接环状GMP和环状di-GMP信号的新型效应子相互作用来调节毒力和生物膜形成。

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