Rhodobacter capsulatus nifA mutants mediating nif gene expression in the presence of ammonium.
Expression of nitrogen fixation genes in Rhodobacter capsulatus is repressed by ammonium at different regulatory levels including an NtrC-independent mechanism controlling NifA activity. In contrast to R. capsulatus NifA, heterologous NifA proteins of Klebsiella pneumoniae and Rhizobium meliloti, respectively, were not subjected to this posttranslational ammonium control in R. capsulatus. The characterization of ammonium-tolerant R. capsulatus NifA1 mutants indicated that the N-terminal domain of NifA was involved in posttranslational regulation. Analysis of a double mutant carrying amino acid substitutions in both the N-terminal domain and the C-terminal DNA-binding domain gave rise to the hypothesis that an interaction between these two domains might be involved in ammonium regulation of NifA activity. Western analysis demonstrated that both constitutively expressed wild-type and ammonium-tolerant NifA1 proteins exhibited high stability and accumulated to comparable levels in cells grown in the presence of ammonium excluding the possibility that proteolytic degradation was responsible for ammonium-dependent inactivation of NifA.
Paschen
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Fakultät für Biologie, Lehrstuhl für Biologie der Mikroorganismen, Ruhr-Universität Bochum, D-44780, Bochum, Germany.
Drepper
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T
Masepohl
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Klipp
W
W
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D017353:Gene Deletion; D015870:Gene Expression / Q000187:drug effects*; D005816:Genetic Complementation Test; D016296:Mutagenesis; D009586:Nitrogen Fixation / Q000187:drug effects; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology*; D016354:Rhodobacter capsulatus / Q000187:drug effects* / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D015533:Transcriptional Activation / Q000187:drug effects
D001426:Bacterial Proteins; C061338:NifA protein, Bacteria; D000644:Quaternary Ammonium Compounds; D014157:Transcription Factors