Frameshifts close to the Klebsiella pneumoniae nifH promoter prevent multicopy inhibition by hybrid nifH plasmids

Abstract

Certain multicopy plasmids bearing promoter sequences of nitrogen fixation (nif) genes inhibit expression of chromosomal genes in Nif⁺ Klebsiella pneumoniae, hence leading to a Nif^- phenotype. This ‘multicopy inhibition’ has been attributed to the titration of the nif-specific activator protein NifA by the plasmid-borne promoter sequences. We now report that multicopy inhibition by nifH translational fusions is sensitive to frameshifts close to the nifH promoter. Transcriptional nifH fusion plasmids in which translation terminated near the nifH promoter were transcriptionally active and showed multicopy inhibition; introduction of a transcription terminator after the nifH coding sequence in these plasmids prevented their multicopy inhibition. Therefore it seems likely that premature termination of transcription prevents multicopy inhibition by the nifH promoter.