Gene fusions were used to demonstrate complex overlapping transcriptional controls of the regulatory gene nifA in Rhizobium meliloti bacteroids isolated from alfalfa root nodules. Gene nifA has previously been shown to be transcribed from promoter PnifA and to be required for activation of promoters P1 (nifHDK) and P2 (fixABC) during symbiotic nitrogen fixation with alfalfa. P2 is located approximately 4 kb upstream from nifA and is shown in this report to be responsible for at least one-half of the nifA expression observed in bacteroids. Substantial transcription of nifA occurs during symbiosis as evidenced by the fact that PnifA was found to be 53% as active as either P1 or P2. Together, the data indicate that more than half of the transcripts initiated at P2 fail to terminate before reaching nifA. Additional studies indicated that there may be weak promoter activity in the symbiotically essential region downstream from nifA.