The nodD protein does not bind to the promoters of inducible nodulation genes in extracts of bacteroids of Rhizobium leguminosarum biovar viciae

In a previous study, we showed that in bacteroids, transcription of the inducible nod genes does not occur and expression of nodD is decreased by 65% (H. R. M. Schlaman, B. Horvath, E. Vijgenboom, R. J. H. Okker, and B. J. J. Lugtenberg, J. Bacteriol. 173:4277-4287, 1991). In the present study, we show, using gel retardation, that in crude extracts of bacteroids of Rhizobium leguminosarum biovar (bv.) viciae, NodD protein does not bind to the nodF, nodM, and nodO box and that it binds only weakly to the nodA box. Binding of NodD from bacteroids to nod box DNA could be restored by mild proteinase K treatment, indicating that NodD is present in bacteroids in an altered form or complex which prevents its binding to nod box DNA. In addition, a novel nodA box DNA-protein complex was found which is specific for the nodA promoter region. This novel complex was formed neither with material from cultured bacterial cells nor with an extract from uninfected roots, and it did not contain NodD but another protein. These results are consistent with the hypothesis that the protein present in the novel retardation complex acts as a transcriptional repressor causing the decreased nodD expression in bacteroids. Such a repressor also explains the lack of NodABCIJ transcription despite the weak NodD binding to the nodA box.