The C-terminal receiver domain of the Rhizobium leguminosarum bv. viciae FixL protein is required for free-living microaerobic induction of the fnrN promoter
The Rhizobium leguminosarum bv. viciae VF39 FixL protein belongs to a distinct group of hybrid regulatory sensor proteins that bear a covalently linked C-terminal receiver domain. FixL has an unorthodox histidine kinase domain, which is shared with many other hybrid regulators. The purified FixL protein had autophosphorylation activity. A truncated protein, lacking the receiver domain, had a much-reduced autophosphorylation activity. However, this truncated protein still efficiently phosphorylated the purified receiver domain in trans. This indicates that, in the full-length FixL protein, the conserved histidine residue in the kinase domain is phosphorylated only transiently and that most of the phosphoryl label accumulates in the C-terminal receiver domain. Gene-fusion studies showed that the fixL gene is required for free-living microaerobic induction of the fnrN promoter. The presence of a functional fixK gene is not required. An R. leguminosarum strain lacking fixL could not be complemented with a truncated copy of the gene lacking the receiver domain. This indicates that the C-terminal receiver domain is an intermediate in the signal transduction pathway that links oxygen limitation to induction of the fnrN promoter in R. leguminosarum bv. viciae VF39.
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The C-terminal receiver domain of the Rhizobium leguminosarum bv. viciae FixL protein is required for free-living microaerobic induction of the fnrN promoter