Genetic analysis reveals links between lipid A structure and expression of the outer membrane protein gene, ropB, in Rhizobium leguminosarum.

The fabXL genes encode enzymes that synthesize the very-long-chain fatty acid - a unique acyl modification located at the 2' position of the lipid A of Gram-negative bacteria in the order Rhizobiales. Mutation of the fabXL genes causes sensitivity to outer membrane stressors and other envelope-related stresses; however, the underlying mechanisms for increased sensitivity are poorly understood. We found that expression of the outer membrane protein gene ropB is down-regulated in an acpXL mutant. Furthermore, constitutive expression of ropB in an acpXL or fabF2XL, fabF1XL mutant restores tolerance to detergents, hyperosmotic stress, and acidic pH. The fabF2XL, fabF1XL mutant also has a delayed nodulation phenotype, whereas a ropB mutant has no observable defects in nodulation, demonstrating that mutation of the fabXL genes results in pleiotropic phenotypes that can be classified as either ropB dependent or ropB independent. Ex-nodule isolates of the mutant strains display restored tolerance to detergents and hyperosmotic and acidic stress conditions; however, the rescued phenotypes are not owing to increased ropB expression. Finally, we found that the fabXL genes are induced by the sensor kinase ChvG in response to peptide-rich growth conditions, which is similar to the results reported for induction of ropB.