An Adenosine Kinase Exists in Xanthomonas campestris Pathovar campestris and Is Involved in Extracellular Polysaccharide Production, Cell Motility, and Virulence

An Adenosine Kinase Exists in Xanthomonas campestris Pathovar campestris and Is Involved in Extracellular Polysaccharide Production, Cell Motility, and Virulence
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腺苷激酶存在于野油菜黄单胞菌中,参与细胞外多糖的产生、细胞运动和毒力

DOI:
10.1128/jb.00009-09
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发表时间:
2009-03
影响因子:
3.2
通讯作者:
陆光涛
陆光涛
中科院分区:
生物学3区
文献类型:
--
作者:
陆光涛

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摘要 腺苷激酶(ADK)是一种嘌呤补救酶,也是真核生物中典型的看家酶,可催化腺苷磷酸化形成 AMP。由于原核生物从头合成嘌呤,并且在大肠杆菌中检测不到内源性 ADK 活性,因此长期以来 ADK 被认为在细菌中很少见。迄今为止,只有两种原核生物(均为革兰氏阳性菌)被报道含有 ADK。在这里,我们报道了革兰氏阴性细菌野油菜黄单胞菌(Xanthomonascampestrispathovarcampestris)(十字花科植物黑腐病的致病菌)拥有编码ADK(名为ADKXcc)的基因(指定为adkXcc),并且我们从遗传学角度证明ADKXcc参与细胞外多糖(EPS)的产生、细胞运动和野油菜黄单胞菌的致病性。 光伏。油菜。 adk Xcc 在大肠杆菌中过表达为 His6 标签蛋白,纯化的 His6 标签蛋白表现出 ADK 活性。 adk Xcc 的突变不会影响细菌在丰富培养基和基本培养基中的生长,但会导致细胞内腺苷的积累以及细胞内 ADK 活性和 ATP 水平以及 EPS 的降低。 adk Xcc 突变体表现出宿主植物中细菌生长和毒力的显着减少。
ABSTRACT Adenosine kinase (ADK) is a purine salvage enzyme and a typical housekeeping enzyme in eukaryotes which catalyzes the phosphorylation of adenosine to form AMP. Since prokaryotes synthesize purines de novo and no endogenous ADK activity is detectable in Escherichia coli, ADK has long been considered to be rare in bacteria. To date, only two prokaryotes, both of which are gram-positive bacteria, have been reported to contain ADK. Here we report that the gram-negative bacterium Xanthomonas campestris pathovar campestris, the causal agent of black rot of crucifers, possesses a gene (designated adk Xcc) encoding an ADK (named ADKXcc), and we demonstrate genetically that the ADKXcc is involved in extracellular polysaccharide (EPS) production, cell motility, and pathogenicity of X. campestris pv. campestris. adk Xcc was overexpressed as a His6-tagged protein in E. coli, and the purified His6-tagged protein exhibited ADK activity. Mutation of adk Xcc did not affect bacterial growth in rich and minimal media but led to an accumulation of intracellular adenosine and diminutions of intracellular ADK activity and ATP level, as well as EPS. The adk Xcc mutant displayed significant reductions in bacterial growth and virulence in the host plant.
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