Mutations in β-tubulin map to domains involved in regulation of microtubule stability in epothilone-resistant cell lines

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Abstract

The epothilones (Epos) are a group of natural products isolated from the myxobacterium, Sorangium cellulosum. They have a mechanism of action similar to that of Taxol, i.e., they stabilize microtubules and induce the formation of microtubule bundles in cells. Because they are simpler in structure than Taxol and preserve their activity in P-glycoprotein-expressing cells, they are being studied as potential antitumor drugs. In this work, a series of Epo-resistant A549 and HeLa cell lines have been selected and analyzed. Class I β-tubulin, the major isotype of β-tubulin in these Epo-resistant cell lines, has been sequenced in a search for mutations. In the Epo B-resistant A549 cells, there is a mutation at β292 from Gln to Glu, in the Epo A-resistant HeLa cell line there is a mutation at β173 from Pro to Ala, and in the Epo B-resistant HeLa cell line there is a heterozygous mutation at β422 from Tyr to a mixture of Tyr and Cys. These mutations are close to the M-loop, the nucleotide-binding site, and the microtubule-associated protein binding sites, respectively. It is likely that these mutations in β-tubulin provide cells with a mechanism of resistance to the Epos and taxanes. Among these resistant cell lines, A549.EpoB40 is hypersensitive to microtubule-destabilizing drugs, such as vinblastine and colchicine, and HeLa.EpoB1.8 is dependent on the Epos or taxanes for growth. Our studies provide evidence that the M-loop, the GTP binding site, and the microtubule-associated protein binding sites at the COOH terminus in β-tubulin are critical for the regulation of microtubule stability.

Original languageEnglish (US)
Pages (from-to)3-10
Number of pages8
JournalMolecular Cancer Therapeutics
Volume1
Issue number1
StatePublished - Nov 2001

Fingerprint

Epothilones
Tubulin
Microtubules
Cell Line
Mutation
Binding Sites
HeLa Cells
Taxoids
Microtubule-Associated Proteins
Paclitaxel
Protein Binding
Myxococcales
Vinblastine
Colchicine
P-Glycoprotein
Guanosine Triphosphate
Biological Products
Antineoplastic Agents
Nucleotides
Growth

ASJC Scopus subject areas

  • Oncology
  • Cancer Research
  • Drug Discovery
  • Pharmacology

Cite this

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title = "Mutations in β-tubulin map to domains involved in regulation of microtubule stability in epothilone-resistant cell lines",
abstract = "The epothilones (Epos) are a group of natural products isolated from the myxobacterium, Sorangium cellulosum. They have a mechanism of action similar to that of Taxol, i.e., they stabilize microtubules and induce the formation of microtubule bundles in cells. Because they are simpler in structure than Taxol and preserve their activity in P-glycoprotein-expressing cells, they are being studied as potential antitumor drugs. In this work, a series of Epo-resistant A549 and HeLa cell lines have been selected and analyzed. Class I β-tubulin, the major isotype of β-tubulin in these Epo-resistant cell lines, has been sequenced in a search for mutations. In the Epo B-resistant A549 cells, there is a mutation at β292 from Gln to Glu, in the Epo A-resistant HeLa cell line there is a mutation at β173 from Pro to Ala, and in the Epo B-resistant HeLa cell line there is a heterozygous mutation at β422 from Tyr to a mixture of Tyr and Cys. These mutations are close to the M-loop, the nucleotide-binding site, and the microtubule-associated protein binding sites, respectively. It is likely that these mutations in β-tubulin provide cells with a mechanism of resistance to the Epos and taxanes. Among these resistant cell lines, A549.EpoB40 is hypersensitive to microtubule-destabilizing drugs, such as vinblastine and colchicine, and HeLa.EpoB1.8 is dependent on the Epos or taxanes for growth. Our studies provide evidence that the M-loop, the GTP binding site, and the microtubule-associated protein binding sites at the COOH terminus in β-tubulin are critical for the regulation of microtubule stability.",
author = "Lifeng He and Yang, {Chia-Ping H.} and {Band Horwitz}, Susan",
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N2 - The epothilones (Epos) are a group of natural products isolated from the myxobacterium, Sorangium cellulosum. They have a mechanism of action similar to that of Taxol, i.e., they stabilize microtubules and induce the formation of microtubule bundles in cells. Because they are simpler in structure than Taxol and preserve their activity in P-glycoprotein-expressing cells, they are being studied as potential antitumor drugs. In this work, a series of Epo-resistant A549 and HeLa cell lines have been selected and analyzed. Class I β-tubulin, the major isotype of β-tubulin in these Epo-resistant cell lines, has been sequenced in a search for mutations. In the Epo B-resistant A549 cells, there is a mutation at β292 from Gln to Glu, in the Epo A-resistant HeLa cell line there is a mutation at β173 from Pro to Ala, and in the Epo B-resistant HeLa cell line there is a heterozygous mutation at β422 from Tyr to a mixture of Tyr and Cys. These mutations are close to the M-loop, the nucleotide-binding site, and the microtubule-associated protein binding sites, respectively. It is likely that these mutations in β-tubulin provide cells with a mechanism of resistance to the Epos and taxanes. Among these resistant cell lines, A549.EpoB40 is hypersensitive to microtubule-destabilizing drugs, such as vinblastine and colchicine, and HeLa.EpoB1.8 is dependent on the Epos or taxanes for growth. Our studies provide evidence that the M-loop, the GTP binding site, and the microtubule-associated protein binding sites at the COOH terminus in β-tubulin are critical for the regulation of microtubule stability.

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