Isolation and characterization of strains defective in vacuolar ornithine permease in Neurospora crassa.
Vacuolar uptake of ornithine and lysine was characterized in Neurospora crassa using a cupric ion permeabilization system. Michaelis constants were measured as 1.4 mM for lysine and 11.0 mM for ornithine, and maximal velocities were determined. Vacuolar lysine uptake was shown to be inhibited competitively by L-arginine and histidine while ornithine uptake was inhibited by a variety of amino acids. Strains defective in the vacuolar ornithine permease were isolated using a filtration enrichment method. Two isolates--RSC-39 and RSC-63--had a reduced ability to accumulate ornithine. Vacuolar uptake of amino acids was measured using cupric ion-permeabilized mycelia; both strains had reduced ornithine uptake while lysine uptake and arginine uptake were normal. For both isolates, both the Michaelis constant and the maximal velocity for ornithine uptake were reduced compared to those of wild type. These results suggest that both strains are defective in the gene which encodes the vacuolar ornithine permease.
Keenan
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KA
Natural and Mathematical Sciences, Richard Stockton College of New Jersey, Pomona 08240, USA. Keenank@pollux.stockton.edu
Utzat
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CD
Zielinski
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TK
D026905:Amino Acid Transport Systems; D001120:Arginine / Q000378:metabolism / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects; D003300:Copper / Q000494:pharmacology; D005800:Genes, Fungal; D006639:Histidine / Q000494:pharmacology; D007700:Kinetics; D008239:Lysine / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D009492:Neurospora crassa / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D009952:Ornithine / Q000378:metabolism*; D013379:Substrate Specificity; D014617:Vacuoles / Q000201:enzymology*
D026905:Amino Acid Transport Systems; D026901:Membrane Transport Proteins; D006639:Histidine; D003300:Copper; D001120:Arginine; D009952:Ornithine; D008239:Lysine