Genetic and biochemical characterization of glutamine synthetase from Neurospora crassa glutamine auxotrophs and their revertants

Abstract

In this paper we present the isolation and characterization of glutamine auxotrophs of Neurospora crassa and their revertants. The results show that although various enrichment procedures were used, we found only two types of auxotrophs. Genetic crosses performed between the different mutants showed that the mutations responsible for their phenotypes were highly linked and probably affected the same gene. The biochemical characterization of the glutamine synthetase polypeptides of the different mutants showed that both types contained the alpha monomer. However, in place of the normal beta monomer, each type had a new polypeptide differing from normal beta either in its molecular weight or in its isoelectric point. On the other hand, the revertants had only the alpha monomer and were capable of growing without glutamine. On the basis of these data, we propose that the lack of glutamine synthetase activity in the auxotrophs is due to the interaction of the altered beta with the alpha monomer, and as a consequence the alpha monomer of the revertants regains its activity because of the absence of the altered beta.