We have focused our attention on arginine metabolism in Neurospora crassa because it encompasses many of the compartmental features characteristic of eukaryotic cells. Biosynthesis of arginine originates in the mitochondria but culminates in the cytosol. Intermediates and arginine cross both mitochondrial membranes. More than 95% of the intramycelial pool of arginine is sequestered in the vacuoles. Our hypothesis is that these compartmentation features play a significant role in the biology of the organism.
We are investigating the metabolic consequences of relocating the arginine biosynthetic enzymes from the mitochondrial matrix to the cytosol. In addition, we are examining how variations in cytosolic arginine concentrations are communicated across the mitochondrial membranes to coordinately inhibit two enzymes of arginine biosynthesis. Molecular techniques are being used to construct mutants defective in the movement of metabolites across intracellular membranes. Control of arginine degradation is being examined by characterizing the expression, structure and properties of arginase, the initial catabolic enzyme. The results of these experiments will provide insight into the function of enzyme and amino acid compartmentation in eukaryotic cells.