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Src family kinases mediate cytoplasmic retention of activated STAT5 in BCR–ABL-positive cells

Oncogene volume 32, pages 3587–3597 (2013)Cite this article

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Abstract

Persistent activation of the Abl tyrosine kinase in the BCR–ABL fusion protein is the major cause of chronic myeloid leukemia (CML). Among many other substrates BCR–ABL phosphorylates STAT5 and Src family kinases (SFK). Activated pSTAT5 is essential for initial transformation and maintenance of the disease. Cytokine-induced phosphorylation on tyrosine 694 typically leads to nuclear accumulation of pSTAT5 and target gene expression. We verified that in BCR–ABL-positive progenitor cells from a CML patient and in K562 cells pSTAT5 is cytoplasmic. However, upon ectopic expression of BCR–ABL p210 in non-myeloid cells, co-transfected STAT5A is phosphorylated on Y694 and localized in the nucleus arguing for an additional factor mediating cytoplasmic retention in CML cells. Expression of the SFK v-Src, Hck or Lyn together with STAT5A results in phosphorylation on Y694 and cytoplasmic retention. Upon coexpression of BCR–ABL and individual SFK the cytoplasmic retention of activated STAT5A mediated by v-Src and Hck but not Lyn is dominant over nuclear translocation induced by BCR–ABL. Cytoplasmic retention depends on the kinase activity of SFK and is mediated through the interaction of the SH2 domain of STAT5A with the SFK. Interestingly, nuclear accumulation of STAT5A as a result of activation by FLT3-ITD, an oncogene found in acute myeloid leukemia, cannot be prevented by coexpression of SFK. Importantly, inhibition of SFK in K562 cells restored nuclear accumulation of pSTAT5A, enhanced STAT5 target gene expression and increased colony formation. Thus, SFK mediate cytoplasmic retention of pSTAT5A in BCR–ABL-positive cells. Cytoplasmic pSTAT5A in CML cells might balance the controversial functions of STAT5 in cellular senescence and differentiation versus G1/S progression and survival.

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Acknowledgements

The research project was supported by grants from the Deutsche Forschungsgemeinschaft (DFG, SFB 542, projects B12 and Z1 to GMN), the Austrian Science Funds (FWF, SFB F28 to RM) and the core facility confocal microscopy of the IZKF Aachen (Interdisciplinary Center for Clinical Research within the Faculty of Medicine at RWTH Aachen University). We would like to thank Frank-D Böhmer, Tim Brümmendorf, Michael Huber, Isabelle Maridonneau-Parini, Fred Schaper and Veronika Sexl for their critical discussion and helping us with plasmid constructs.

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Authors and Affiliations

  1. Institut für Biochemie und Molekularbiologie, RWTH Aachen University, Aachen, Germany

    N Chatain, D Fahrenkamp, H Schmitz-Van de Leur & G Müller-Newen

  2. Klinik für Onkologie, Hämatologie und Stammzelltransplantation, RWTH Aachen University, Aachen, Germany

    P Ziegler & E Jost

  3. Ludwig Boltzmann Institute for Cancer Research (LBI-CR), Vienna, Austria

    R Moriggl

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Chatain, N., Ziegler, P., Fahrenkamp, D. et al. Src family kinases mediate cytoplasmic retention of activated STAT5 in BCR–ABL-positive cells. Oncogene 32, 3587–3597 (2013). https://doi.org/10.1038/onc.2012.369

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  • DOI: https://doi.org/10.1038/onc.2012.369

Keywords

  • CML
  • BCR–ABL
  • STAT5
  • Src
  • inhibitors

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