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Phosphorylation of tyrosine 393 in the kinase domain of Bcr-Abl influences the sensitivity towards imatinib in vivo

Leukemia volume 17, pages 1695–1699 (2003)Cite this article

Abstract

The Bcr-Abl fusion protein arising through the t(9;22)(q34;q11) reciprocal translocation is the causative agent in chronic myeloid leukemia and a subset of acute lymphocytic leukemia. Imatinib mesylate is a specific inhibitor of the Bcr-Abl kinase and has shown promising results in clinical studies. The structural relation between the Bcr-Abl oncogene and the tyrosine kinase inhibitor imatinib has recently been elucidated by an elegant crystal structure analysis, emphasizing the importance of dephosphorylated tyrosine 393 (Tyr393) in Bcr-Abl for access of the inhibitor to the kinase domain. By mutating this tyrosine to phenylalanine and thereby mimicking a constitutively dephosphorylated state, we now show that Ba/F3 cells transformed by this mutant demonstrate an increased sensitivity towards imatinib in vivo. This effect is not due to an impaired kinase activity of Bcr-Abl Y393F, since a synthetic substrate is phosphorylated with similar kinetics. Treatment of Ba/F3 cells transfected with Bcr-Abl wild type with a phosphatase inhibitor diminished the effect of imatinib, but did not influence the growth of Ba/F3 cells transfected with Bcr-AblY393F. The results support the findings of the crystal structure and indicate that Tyr393 indeed plays a significant role for the sensitivity of Bcr-Abl towards imatinib in vivo. These data implicate the regulation of Tyr393 phosphorylation as a potential mechanism of imatinib resistance.

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Acknowledgements

We are gratefully indebted to Dr H Gschaidmeier (Novartis Pharma, Nürnberg, Germany) and Dr E Buchdunger (Novartis Pharma, Basel, Switzerland) for providing imatinib and to Dr T Wheeler (Dairy Science, AgResearch, Ruakura Research Centre, Hamilton, New Zealand) for the phospho-STAT5 antibody.

This work was supported by a grant to JD and CP from the BMBF, German national genome Project No. 01-GS-0105 and 01-GS-0155 and by a grant from the Mildred-Scheel Stiftung to JD. CM is supported by a fellowship from the Deutsche Jose Carreras Leukämie Stiftung (DJCLS 2001/NAT-2).

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  1. Department of Internal Medicine III, Technical University of Munich, Munich, Germany

    C Miething, C Mugler, R Grundler, J Hoepfl, R-Y Bai, C Peschel & J Duyster

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  2. C Mugler
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Miething, C., Mugler, C., Grundler, R. et al. Phosphorylation of tyrosine 393 in the kinase domain of Bcr-Abl influences the sensitivity towards imatinib in vivo. Leukemia 17, 1695–1699 (2003). https://doi.org/10.1038/sj.leu.2403040

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