THE INTEGRATION HOST FACTOR STIMULATES INTERACTION OF RNA-POLYMERASE WITH NIFA, THE TRANSCRIPTIONAL ACTIVATOR FOR NITROGEN-FIXATION OPERONS

The regulatory protein NIFA activates transcription of nitrogen fixation (nif) operons by the σ54 holoenzyme form of RNA polymerase. NIFA from Klebsiella pneumoniae activates transcription from the nifH promoter in vitro; in addition, the integration host factor, IHF, binds between the nifH promoter and an upstream binding site for NIFA. We demonstrate here that IHF greatly stimulates NIFA-mediated activation of nifH transcription in vitro and thus that the two factors are functionally synergistic. Electron micrographs indicate that IHF bends the DNA in the nifH promoter regulatory region. Although IHF binds close to the nifH promoter, it does not directly stlmulate binding of σ54 holoenzyme. Rather, the IHF-induced bend may facilitate productive contacts between NIFA and σ54 holoenzyme that lead to the formation of open complexes. IHF binds to nif promoter regulatory regions from a variety of organisms within the phylum "purple bacteria," suggesting a general ability to stimulate NIFA-mediated activation of nif transcription. © 1990.