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Elucidation of protein interactions necessary for the maintenance of the BCR–ABL signaling complex

Abstract

Many patients with chronic myeloid leukemia in deep remission experience return of clinical disease after withdrawal of tyrosine kinase inhibitors (TKIs). This suggests signaling of inactive BCR–ABL, which allows the survival of cancer cells, and relapse. We show that TKI treatment inhibits catalytic activity of BCR–ABL, but does not dissolve BCR–ABL core signaling complex, consisting of CRKL, SHC1, GRB2, SOS1, cCBL, p85a-PI3K, STS1 and SHIP2. Peptide microarray and co-immunoprecipitation results demonstrate that CRKL binds to proline-rich regions located in C-terminal, intrinsically disordered region of BCR–ABL, that SHC1 requires pleckstrin homology, src homology and tyrosine kinase domains of BCR–ABL for binding, and that BCR–ABL sequence motif located in disordered region around phosphorylated tyrosine 177 mediates binding of three core complex members, i.e., GRB2, SOS1, and cCBL. Further, SHIP2 binds to the src homology and tyrosine kinase domains of BCR–ABL and its inositol phosphatase activity contributes to BCR–ABL-mediated phosphorylation of SHC1. Together, this study characterizes protein–protein interactions within the BCR–ABL core complex and determines the contribution of particular BCR–ABL domains to downstream signaling. Understanding the structure and dynamics of BCR–ABL interactome is critical for the development of drugs targeting integrity of the BCR–ABL core complex.

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Acknowledgements

This study was supported by Agency for Healthcare Research of the Czech Republic, project NV15-34405A. AN was supported by European Regional Development Fund OP RDE Project no. CZ.02.1.01/0.0/0.0/16_019/0000729.

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Correspondence to Lukas Trantirek or Pavel Krejci.

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Gregor, T., Bosakova, M.K., Nita, A. et al. Elucidation of protein interactions necessary for the maintenance of the BCR–ABL signaling complex. Cell. Mol. Life Sci. 77, 3885–3903 (2020). https://doi.org/10.1007/s00018-019-03397-7

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Keywords

  • BCR–ABL
  • Chronic myeloid leukemia
  • Signaling
  • Protein complex