Abstract
The Gram-positive soil bacterium Bacillus subtilis is able to choose between motile and sessile lifestyles. The sessile way of life, also referred to as biofilm, depends on the formation of an extracellular polysaccharide matrix and some extracellular proteins. Moreover, a significant proportion of cells in a biofilm form spores. The first two genes of the 15-gene operon for extracellular polysaccharide synthesis, epsA and epsB, encode a putative transmembrane modulator protein and a putative protein tyrosine kinase, respectively, with similarity to the TkmA/PtkA modulator/kinase couple. Here we show that the putative kinase EpsB is required for the formation of structured biofilms. However, an epsB mutant is still able to form biofilms. As shown previously, a ptkA mutant is also partially defective in biofilm formation, but this defect is related to spore formation in the biofilm. The absence of both kinases resulted in a complete loss of biofilm formation. Thus, EpsB and PtkA fulfil complementary functions in biofilm formation. The activity of bacterial protein tyrosine kinases depends on their interaction with modulator proteins. Our results demonstrate the specific interaction between the putative kinase EpsB and its modulator protein EpsA and suggest that EpsB activity is stimulated by its modulator EpsA.
Highlights
Like many other bacteria, the Gram-positive model organism Bacillus subtilis is able to form structured biofilms in order to attach to surfaces and to withstand harsh environmental conditions
Bacillus subtilis cells in the biofilm form an extracellular matrix composed of polysaccharides, amyloid-like fibres made up of the TasA protein, and a water-repellent surface coat formed by the bacterial hydrophobin BslA
The EpsA and EpsB proteins are similar to the TkmA transmembrane kinase modulator and the PtkA protein tyrosine kinase of Bacillus subtilis, respectively
Summary
The Gram-positive model organism Bacillus subtilis is able to form structured biofilms in order to attach to surfaces and to withstand harsh environmental conditions. Biofilm formation in Bacillus subtilis is usually observed as the formation of either structured complex colonies on solid surfaces or thick floating pellicles in liquid media (Branda et al, 2001). Biofilm formation is strongly impaired in the domesticated laboratory strain of Bacillus subtilis, 168. This strain forms only poorly structured colonies and thinner and non-structured yet stable pellicles. Such a loss of phenotype, which is important in natural environments but not in the laboratory, is commonly observed in bacteria. Reduced biofilm formation in the Abbreviations: B2H, bacterial two-hybrid; EPS, exopolysaccharide
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