GlnK effects complex formation between NifA and NifL in Klebsiella pneumoniae.
In Klebsiella pneumoniae, the nif specific transcriptional activator NifA is inhibited by NifL in response to molecular oxygen and ammonium. Here, we demonstrate complex formation between NifL and NifA (approximately 1 : 1 ratio), when synthesized in the presence of oxygen and/or ammonium. Under simultaneous oxygen- and nitrogen-limitation, significant but fewer NifL-NifA complexes (approximately 1%) were formed in the cytoplasm as a majority of NifL was sequestered to the cytoplasmic membrane. These findings indicate that inhibition of NifA in the presence of oxygen and/or ammonium occurs via direct NifL interaction and formation of those inhibitory NifL-NifA complexes appears to be directly and exclusively dependent on the localization of NifL in the cytoplasm. We further observed evidence that the nitrogen sensory protein GlnK forms a trimeric complex with NifL and NifA under nitrogen limitation. Binding of GlnK to NifL-NifA was specific; however the amount of GlnK within these complexes was small. Finally, two lines of evidence were obtained that under anaerobic conditions but in the presence of ammonium additional NtrC-independent GlnK synthesis inhibited the formation of stable inhibitory NifL-NifA complexes. Thus, we propose that the NifL-NifA-GlnK complex reflects a transitional structure and hypothesize that under nitrogen-limitation, GlnK interacts with the inhibitory NifL-NifA complex, resulting in its dissociation.
Stips
Jessica
J
Institut für Mikrobiologie und Genetik, Göttingen, Germany.
Thummer
Robert
R
Neumann
Melanie
M
Schmitz
Ruth A
RA
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D007711:Klebsiella pneumoniae / Q000235:genetics / Q000378:metabolism*; D046911:Macromolecular Substances; D009584:Nitrogen / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D011485:Protein Binding; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D046911:Macromolecular Substances; C061338:NifA protein, Bacteria; D000644:Quaternary Ammonium Compounds; D011994:Recombinant Proteins; D014157:Transcription Factors; C084296:nifL protein, Bacteria; D009584:Nitrogen; D010100:Oxygen