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 sigma 54 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 stimulate binding of sigma 54 holoenzyme. Rather, the IHF-induced bend may facilitate productive contacts between NIFA and sigma 54 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.
Hoover
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TR
Department of Plant Pathology, University of California, Berkeley 94720.
Santero
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Porter
S
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Kustu
S
S
D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D004254:DNA Nucleotidyltransferases / Q000378:metabolism* / Q000648:ultrastructure; D004269:DNA, Bacterial / Q000235:genetics / Q000648:ultrastructure; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D019426:Integrases; D007711:Klebsiella pneumoniae / Q000235:genetics*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009586:Nitrogen Fixation / Q000235:genetics*; D009876:Operon; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D013698:Templates, Genetic; D014157:Transcription Factors; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D004269:DNA, Bacterial; C061338:NifA protein, Bacteria; D014157:Transcription Factors; D004254:DNA Nucleotidyltransferases; D019426:Integrases; D012321:DNA-Directed RNA Polymerases