Positive involvement of ppGpp in derepression of the nif operon in Klebsiella pneumoniae

Abstract

The kinetics of derepression of the enzyme nitrogenase were investigated, after exhaustion of a limiting amount of ammonium from the culture medium, in a prototrophic stringent-relaxed pair of Klebsiella pneumoniae strains and in their F relA+-F relA derivatives. The results indicate that ppGpp (guanosine 3'-5' diphosphate) increases the nitrogen fixation capability of K. pneumoniae by at least three different mechanisms. (1) It prevents exhaustion of the ATP pool when nitrogen starvation is imposed. (2) The translational defects in relaxed mutants are suppressed by ppGpp during nif derepression. (3) The synthesis of nitrogenase components is at least five times higher in the presence of ppGpp than in its absence. This latter conclusion was based on experimental results obtained when following the incorporation of (35S)-methionine into nitrogenase components after pulse labelling at various time intervals during nif derepression. The nitrogenase components were separated by solid phase radioimmunoassay as well as by two-dimensional gel electrophoresis.