Rhizoxin resistant mutants with an altered beta-tubulin gene in Aspergillus nidulans.
Rhizoxin and ansamitocin P-3 (a maytansinoid compound), potent inhibitors of mammalian brain tubulin assembly, inhibit growth of a variety of fungi including Aspergillus nidulans. Mutants of A. nidulans, benA10 which is a benomyl resistant beta-tubulin gene mutant and tubA1 which is a benomyl supersensitive alpha-tubulin gene mutant, were both sensitive to rhizoxin and ansamitocin P-3 to the same extent as wild-type strains. We isolated 18 rhizoxin resistant mutants of A. nidulans. All of these mutants were cross-resistant to ansamitocin P-3, but not to benzimidazole antimitotic drugs. These mutants mapped to two loci, rhiA and rhiB, and all of those with high resistance mapped to rhiA. The fact that the protein extracts of rhiA mutants lost rhizoxin binding affinity and that rhiA was closely linked to benA, the major beta-tubulin gene in A. nidulans, indicated that rhiA must be a structural gene for beta-tubulin and that rhiA mutants are a new class of beta-tubulin gene mutants. All of this suggested that, in A. nidulans, these antimitotic drugs bind to beta-tubulin, and that rhizoxin and ansamitocin P-3 share the same binding site but the site does not overlap with the benzimidazole binding site. Protein extracts from a rhiB mutant retained rhizoxin binding affinity, therefore this rhizoxin resistance mechanism should not be a tubulin mediated process.
Takahashi
M
M
Institute of Applied Microbiology, University of Tokyo, Japan.
Kobayashi
H
H
Iwasaki
S
S
D000935:Antifungal Agents / Q000494:pharmacology*; D001233:Aspergillus nidulans / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D001542:Benomyl / Q000008:administration & dosage / Q000494:pharmacology; D001562:Benzimidazoles / Q000008:administration & dosage / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004352:Drug Resistance, Microbial / Q000235:genetics; D005800:Genes, Fungal; D007783:Lactones / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology; D018942:Macrolides; D009154:Mutation; D014404:Tubulin / Q000235:genetics*
D000935:Antifungal Agents; D001562:Benzimidazoles; D007783:Lactones; D018942:Macrolides; D014404:Tubulin; C041443:rhizoxin; C031000:benzimidazole; D001542:Benomyl