Elevated levels of microtubule destabilizing factors in a Taxol-resistant/dependent A549 cell line with an α-tubulin mutation

Laura A. Martello, Pascal Verdier-Pinard, Heng Jia Shen, Lifeng He, Keila Torres, George A. Orr, Susan Band Horwitz

Research output: Contribution to journalArticle

131 Citations (Scopus)

Abstract

The A549 Taxol-resistant cell lines, A549-T12 and A549-T24, were isolated in our laboratory, and are dependent on Taxol for normal growth. The microtubules in these cells displayed increased dynamicity in the absence of Taxol. In the present study, a heterozygous point mutation in Kα1-tubulin was discovered at α379 (Ser to Ser/Arg). Although Taxol binds to β-tubulin in the microtubule, sequencing of β-tubulin class I did not reveal any mutations. The expression of the α-tubulin mutation was demonstrated using high-resolution isoelectric focusing and two-dimensional gel analysis. Both the wild-type and mutant tubulin were expressed in the Taxol-resistant cell lines. The region of α-tubulin that encompasses α379 is near the COOH terminus that has been proposed as a site of interaction with microtubule-associated protein (MAP) 4 and stathmin, a tubulin-interacting protein. In the Taxol-resistant cells, the active non-phosphorylated form of stathmin was increased ∼2-fold, whereas the inactive phosphorylated forms were barely detected. The inactive phosphorylated forms of MAP4 were increased in the A549-T12 and A549-T24 cell lines. We hypothesize that these changes in tubulin/MAPs that result in increased microtubule instability may be related to the ∩-tubulin mutation and are compensated for by the stabilizing properties of Taxol.

Original languageEnglish (US)
Pages (from-to)1207-1213
Number of pages7
JournalCancer Research
Volume63
Issue number6
StatePublished - Mar 15 2003

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Tubulin
Paclitaxel
Microtubules
Mutation
Stathmin
A549 Cells
Isoelectric Focusing
Point Mutation
Gels
Cell Line
Growth

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Martello, L. A., Verdier-Pinard, P., Shen, H. J., He, L., Torres, K., Orr, G. A., & Band Horwitz, S. (2003). Elevated levels of microtubule destabilizing factors in a Taxol-resistant/dependent A549 cell line with an α-tubulin mutation. Cancer Research, 63(6), 1207-1213.

Elevated levels of microtubule destabilizing factors in a Taxol-resistant/dependent A549 cell line with an α-tubulin mutation. / Martello, Laura A.; Verdier-Pinard, Pascal; Shen, Heng Jia; He, Lifeng; Torres, Keila; Orr, George A.; Band Horwitz, Susan.

In: Cancer Research, Vol. 63, No. 6, 15.03.2003, p. 1207-1213.

Research output: Contribution to journalArticle

Martello, LA, Verdier-Pinard, P, Shen, HJ, He, L, Torres, K, Orr, GA & Band Horwitz, S 2003, 'Elevated levels of microtubule destabilizing factors in a Taxol-resistant/dependent A549 cell line with an α-tubulin mutation', Cancer Research, vol. 63, no. 6, pp. 1207-1213.
Martello LA, Verdier-Pinard P, Shen HJ, He L, Torres K, Orr GA et al. Elevated levels of microtubule destabilizing factors in a Taxol-resistant/dependent A549 cell line with an α-tubulin mutation. Cancer Research. 2003 Mar 15;63(6):1207-1213.
Martello, Laura A. ; Verdier-Pinard, Pascal ; Shen, Heng Jia ; He, Lifeng ; Torres, Keila ; Orr, George A. ; Band Horwitz, Susan. / Elevated levels of microtubule destabilizing factors in a Taxol-resistant/dependent A549 cell line with an α-tubulin mutation. In: Cancer Research. 2003 ; Vol. 63, No. 6. pp. 1207-1213.
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