Conservation between coding and regulatory elements of Rhizobium meliloti and Rhizobium leguminosarum dct genes

Complementation of Rhizobium leguminosarum dct mutants with a cosmid bank yielded Rhizobium meliloti homologs of the dctA, dctB, and dctD genes. The genes dctB and dctD are thought to form a two-component system which responds to the presence of C4-dicarboxylates to regulate expression of a transport protein encoded by dctA. DNA sequence analysis showed that dct coding and intergenic regions, including putative binding sites for the dctD protein and sigma 54-RNA polymerase, were highly conserved between these two Rhizobium species. Mutation of R. meliloti dctD showed that it was not essential for symbiotic nitrogen fixation but was needed for growth on succinate and the expression of a dctA-lacZ fusion gene in free-living cells. Hybridization of R. meliloti genomic DNA with probes representing the central portion of dctD potentially identified more than 20 similar regulatory genes, all of which are likely to depend upon the alternative sigma factor encoded by rpoN and stimulate transcription in a manner very similar to ntrC activation of glnA in enteric bacteria.