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

Abstract

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.