Kinase domain mutants of Bcr-Abl exhibit altered transformation potency, kinase activity, and substrate utilization, irrespective of sensitivity to imatinib

Ian J. Griswold, Mary MacPartlin, Thomas Bumm, Valerie L. Goss, Thomas O'Hare, Kimberly A. Lee, Amie S. Corbin, Eric P. Stoffregen, Caitlyn Smith, Kara Johnson, Erika M. Moseson, Lisa Wood, Roberto D. Polakiewicz, Brian Druker, Michael W. Deininger

Research output: Contribution to journalArticle

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Abstract

Kinase domain (KD) mutations of Bcr-Abl interfering with imatinib binding are the major mechanism of acquired imatinib resistance in patients with Philadelphia chromosome-positive leukemia. Mutations of the ATP binding loop (p-loop) have been associated with a poor prognosis. We compared the transformation potency of five common KD mutants in various biological assays. Relative to unmutated (native) Bcr-Abl, the ATP binding loop mutants Y253F and E255K exhibited increased transformation potency, M351T and H396P were less potent, and the performance of T315I was assay dependent. The transformation potency of Y253F and M351T correlated with intrinsic Bcr-Abl kinase activity, whereas the kinase activity of E255K, H396P, and T315I did not correlate with transforming capabilities, suggesting that additional factors influence transformation potency. Analysis of the phosphotyrosine proteome by mass spectroscopy showed differential phosphorylation among the mutants, a finding consistent with altered substrate specificity and pathway activation. Mutations in the KD of Bcr-Abl influence kinase activity and signaling in a complex fashion, leading to gain- or loss-of-function variants. The drug resistance and transformation potency of mutants may determine the outcome of patients on therapy with Abl kinase inhibitors.

Original languageEnglish (US)
Pages (from-to)6082-6093
Number of pages12
JournalMolecular and Cellular Biology
Volume26
Issue number16
DOIs
StatePublished - Aug 2006

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Phosphotransferases
Mutation
Adenosine Triphosphate
Philadelphia Chromosome
Phosphotyrosine
Proteome
Substrate Specificity
Imatinib Mesylate
Drug Resistance
Biological Assay
Mass Spectrometry
Leukemia
Phosphorylation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

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Kinase domain mutants of Bcr-Abl exhibit altered transformation potency, kinase activity, and substrate utilization, irrespective of sensitivity to imatinib. / Griswold, Ian J.; MacPartlin, Mary; Bumm, Thomas; Goss, Valerie L.; O'Hare, Thomas; Lee, Kimberly A.; Corbin, Amie S.; Stoffregen, Eric P.; Smith, Caitlyn; Johnson, Kara; Moseson, Erika M.; Wood, Lisa; Polakiewicz, Roberto D.; Druker, Brian; Deininger, Michael W.

In: Molecular and Cellular Biology, Vol. 26, No. 16, 08.2006, p. 6082-6093.

Research output: Contribution to journalArticle

Griswold, IJ, MacPartlin, M, Bumm, T, Goss, VL, O'Hare, T, Lee, KA, Corbin, AS, Stoffregen, EP, Smith, C, Johnson, K, Moseson, EM, Wood, L, Polakiewicz, RD, Druker, B & Deininger, MW 2006, 'Kinase domain mutants of Bcr-Abl exhibit altered transformation potency, kinase activity, and substrate utilization, irrespective of sensitivity to imatinib', Molecular and Cellular Biology, vol. 26, no. 16, pp. 6082-6093. https://doi.org/10.1128/MCB.02202-05
Griswold, Ian J. ; MacPartlin, Mary ; Bumm, Thomas ; Goss, Valerie L. ; O'Hare, Thomas ; Lee, Kimberly A. ; Corbin, Amie S. ; Stoffregen, Eric P. ; Smith, Caitlyn ; Johnson, Kara ; Moseson, Erika M. ; Wood, Lisa ; Polakiewicz, Roberto D. ; Druker, Brian ; Deininger, Michael W. / Kinase domain mutants of Bcr-Abl exhibit altered transformation potency, kinase activity, and substrate utilization, irrespective of sensitivity to imatinib. In: Molecular and Cellular Biology. 2006 ; Vol. 26, No. 16. pp. 6082-6093.
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