Enzymology of biological nitrogen fixation

Two genes involved in the regulation of nitrogenase activity, draT and draG, were cloned and found to be contiguous on the Azospirillum brasilense chromosome. The nifH gene, encoding dinitrogenase reductase, is near to draT with an intervening gap of 1.9 kb. The organization of these genes in Azospirillum lipoferum and Rhodosprillum rubrum is similar, but nifH and draT are separated by only 400 bp in the organisms. A. brasilense draTG is very similar to draTG in R. rubrum with 91.8% similarity and 85.3% identity at the amino acid level. Apparently A. brasilense uses the normal ATG initiation codon for draT, and draG. The genes for A. brasilense were able to restore function to appropriate mutants of R. rubrum. The heterologous expression of A. brasilense draTG in R. rubrum was not fully normal, as it responded more slowly to darkness and more quickly to ammonia than wild type cells. Our mutational analysis of the draTG region of A. brasilense confirms the function of these genes in the regulation of nitrogenase activity, but it also revealed minor but demonstrable differences in the control systems of R. rubrum and A. brasilense.