ABSTRACT
Rhizobium leguminosarum is a soil bacterium that infects root hairs and induces the formation of nitrogen-fixing nodules on leguminous plants. Light, oxygen, and voltage (LOV)-domain proteins are blue-light receptors found in higher plants and many algae, fungi, and bacteria. The genome of R. leguminosarum bv. viciae 3841, a pea-nodulating endosymbiont, encodes a sensor histidine kinase containing a LOV domain at the N-terminal end (R-LOV-HK). R-LOV-HK has a typical LOV domain absorption spectrum with broad bands in the blue and UV-A regions and shows a truncated photocycle. Here we show that the R-LOV-HK protein regulates attachment to an abiotic surface and production of flagellar proteins and exopolysaccharide in response to light. Also, illumination of bacterial cultures before inoculation of pea roots increases the number of nodules per plant and the number of intranodular bacteroids. The effects of light on nodulation are dependent on a functional lov gene. The results presented in this work suggest that light, sensed by R-LOV-HK, is an important environmental factor that controls adaptive responses and the symbiotic efficiency of R. leguminosarum.
Subject(s)
Bacterial Adhesion/physiology , Light , Photoreceptors, Microbial/metabolism , Pisum sativum/microbiology , Plant Root Nodulation/physiology , Rhizobium leguminosarum/physiology , Symbiosis , Amino Acid Sequence , Bacterial Adhesion/radiation effects , Base Sequence , Biofilms/growth & development , Blotting, Western , Flagella/metabolism , Gentian Violet , Histidine Kinase , Microscopy, Electron, Scanning , Molecular Sequence Data , Plant Root Nodulation/radiation effects , Polysaccharides, Bacterial/metabolism , Protein Kinases/genetics , Protein Kinases/metabolism , Protein Structure, Tertiary/genetics , Real-Time Polymerase Chain Reaction , Rhizobium leguminosarum/radiation effects , Rhizobium leguminosarum/ultrastructure , Sequence Alignment , Sequence Analysis, DNA , Statistics, NonparametricABSTRACT
Bacteria rely on two-component signaling systems in their adaptive responses to environmental changes. Typically, the two-component system consists of a sensory histidine kinase that signals by transferring a phosphoryl group to a secondary response regulator that ultimately relays the signal to the cell. Some of these sensors use PAS (Per-Arnt-Sin) domains. A new member of the PAS super family is the LOV (light, oxygen, voltage) domain, a 10-kDa flavoprotein that functions as a light-sensory module in plant, algal, fungal, and bacterial blue-light receptors. Putative LOV domains have been identified in the genomes of many higher and lower eukaryotes, plants, eubacteria, archaebacteria, and particularly in genes coding for histidine kinases (LOV-histidine kinases, LOV-HKs) of plant and animal pathogenic bacteria, including Brucella. We describe here biochemical, photochemical, and biophysical methodology to purify these enzymes and to characterize their light-activation process.
Subject(s)
Bacterial Proteins/metabolism , Light , Protein Kinases/metabolism , Bacterial Proteins/genetics , Brucella/enzymology , Brucella/radiation effects , Enzyme Activation/radiation effects , Histidine Kinase , Photochemistry , Protein Kinases/geneticsABSTRACT
Histidine kinases, used for environmental sensing by bacterial two-component systems, are involved in regulation of bacterial gene expression, chemotaxis, phototaxis, and virulence. Flavin-containing domains function as light-sensory modules in plant and algal phototropins and in fungal blue-light receptors. We have discovered that the prokaryotes Brucella melitensis, Brucella abortus, Erythrobacter litoralis, and Pseudomonas syringae contain light-activated histidine kinases that bind a flavin chromophore and undergo photochemistry indicative of cysteinyl-flavin adduct formation. Infection of macrophages by B. abortus was stimulated by light in the wild type but was limited in photochemically inactive and null mutants, indicating that the flavin-containing histidine kinase functions as a photoreceptor regulating B. abortus virulence.
