title: Antibiotic-induced derepression of the NAD-specific glutamate dehydrogenase of Neurospora crassa.
abstract: The catabolic, NAD-specific glutamate dehydrogenase (NAD-GDH) of Neurospora crassa is under carbon catabolite repression. Cells grown on a glycolytic carbon source, such as sucrose, have low basal levels of enzyme activity. Treatment of repressed cells with either polymyxin B or amphotericin B resulted in derepression of NAD-GDH. Derepression at the transcriptional level occurred very rapidly (within 30 min) in response to polymyxin B addition but reached a plateau within 2 h. Amphotericin B-induced derepression initiated more slowly but continued for at least 6 h, resulting in a specific activity comparable to that seen with cells transferred to glutamate as the sole carbon source. These antibiotics had no significant effect upon the activities of two constitutive enzymes, pyruvate kinase and malate dehydrogenase. Curiously, only polymyxin B treatment derepressed invertase, another catabolite-repressed enzyme. The addition of 100 mM KCl to the growth medium blocked derepression by both antibiotics, but the addition of 50 mM MgCl2 only annulled derepression by polymyxin B. The ergosterol-deficient erg-1 mutant, which is resistant to amphotericin B, did not derepress NAD-GDH when treated with this drug. These results are consistent with derepression resulting from interactions of these antibiotics with the plasma membrane.
authors.affiliation: Department of Biological Sciences, University of Calgary, Alberta, Canada.
authors.forename: P J
authors.initials: PJ
authors.lastname: Vierula
authors.forename: M
authors.initials: M
authors.lastname: Kapoor
mesh_terms: D000666:Amphotericin B / Q000494:pharmacology*; D002462:Cell Membrane / Q000201:enzymology; D004794:Enzyme Repression; D005969:Glutamate Dehydrogenase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D009243:NAD / Q000378:metabolism; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D011112:Polymyxin B / Q000494:pharmacology*; D011113:Polymyxins / Q000494:pharmacology*; D014176:Protein Biosynthesis
chemical_list: D011113:Polymyxins; D009243:NAD; D000666:Amphotericin B; D005969:Glutamate Dehydrogenase; D011112:Polymyxin B