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J Bacteriol ; 191(11): 3639-48, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19329636

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 ADK(Xcc)), and we demonstrate genetically that the ADK(Xcc) is involved in extracellular polysaccharide (EPS) production, cell motility, and pathogenicity of X. campestris pv. campestris. adk(Xcc) was overexpressed as a His(6)-tagged protein in E. coli, and the purified His(6)-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.


Subject(s)
Adenosine Kinase/physiology , Bacterial Proteins/physiology , Polysaccharides, Bacterial/biosynthesis , Virulence/genetics , Xanthomonas/enzymology , Xanthomonas/pathogenicity , Adenosine Kinase/genetics , Adenosine Kinase/metabolism , Adenosine Triphosphate/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Mutation , Polysaccharides, Bacterial/genetics , Xanthomonas/genetics
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