Effects of ribonuclease A on amino acid transport in Neurospora crassa.
Incubation of Neurospora crassa conidia with ribonuclease (RNase) A reduces transport of L-phenylalanine by those cells. Under similar conditions, oxidized RNase A, RNase T1, and RNase T2 do not have this effect. Incubation of conidia with active RNase covalently attached to polyacrylamide beads reduces L-phenylalanine transport. This indicates that the site of enzymatic action is at the cell surface. At the lower concentration of enzyme used in this study, incubation with RNase A reduces transport of L-phenylalanine by the general (G) amino acid permease. Increasing the enzyme concentration results in reduction of transport by the neutral aromatic (N)-specific permease. The increased transport activity that accompanies onset of conidial germination is also sensitive to incubation with RNase A. Application of the enzyme to actively transporting cells does not release amino acid transported prior to enzyme addition. Cells cultured on media supplemented with [2-14C] uridine release isotopic activity after RNase A incubation. Analogous treatments with Pronase, RNase T1, RNase T2, or deoxyribonuclease I do not release isotope activity. Pronase treatment does reduce L-phenylalanine transport. Incubation of conidia with RNase A also inhibits germination of those conidia.
D000596:Amino Acids / Q000378:metabolism*; D001693:Biological Transport, Active / Q000187:drug effects; D026901:Membrane Transport Proteins / Q000378:metabolism; D009154:Mutation; D009491:Neurospora / Q000378:metabolism*; D009492:Neurospora crassa / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D010649:Phenylalanine / Q000378:metabolism; D011402:Pronase / Q000494:pharmacology; D012260:Ribonucleases / Q000494:pharmacology*; D013172:Spores, Fungal / Q000254:growth & development / Q000378:metabolism; D014529:Uridine / Q000378:metabolism
D000596:Amino Acids; D026901:Membrane Transport Proteins; D010649:Phenylalanine; D012260:Ribonucleases; D011402:Pronase; D014529:Uridine