Summary
Rhizoxin, an antibiotic, exhibits potent anti-mitotic activity against most eucaryotic cells including those of higher vertebrates, plants and fungi by binding to β-tubulin. ThebenA gene of three independently isolated rhizoxin-resistant (Rhir) mutants ofAspergillus nidulans was cloned, sequenced and compared with that of the wild-type, rhizoxin-sensitive (Rhis) strain. In all three Rhir mutants, the AAC codon for Asn-100 of thebenA β-tubulin gene was altered to ATC, coding for Ile. Sequence displacement experiments confirmed that the substitution of Ile for Asn-100 confers resistance to rhizoxin in this organism. The amino acid sequences of β-tubulin surrounding the 100th amino acid residue from the N-terminus including Asn-100 are highly conserved with a few exceptions. The fission yeastSchizosaccharomyces pombe and the budding yeastSaccharomyces cerevisiae are naturally occurring Rhir organisms whose β-tubulin genes encode Ile and Val respectively at the 100th amino acid residue. The Ile-100 ofS. pombe and the Val-100 ofS. cerevisiae were altered to Asn using site-directed mutagenesis and gene displacement techniques. The resultant haploid strains of these two yeasts uniquely expressing β-tubulin (Asn-100) instead of β-tubulin (Ile-100 or Val-100) were found to be Rhis. Haploid yeast expressing β-tubulin (Asn-100) is normal except for its sensitivity to rhizoxin. These results suggest that rhizoxin resistance has a common basis in both naturally occurring species and experimentally selected mutants in the substitution of Ile or Val for Asn-100 in β-tubulin.
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Takahashi, M., Matsumoto, S., Iwasaki, S. et al. Molecular basis for determining the sensitivity of eucaryotes to the antimitotic drug rhizoxin. Mol Gen Genet 222, 169–175 (1990). https://doi.org/10.1007/BF00633814
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DOI: https://doi.org/10.1007/BF00633814