Key Points
-
Abl is a non-receptor tyrosine kinase that contains a Src-homology-3 (SH3)- and a Src-homology-2 (SH2)-domain. Abl participates in many signalling pathways in the cytoplasm and the nucleus. The oncogenic fusion Bcr–Abl, which is caused by reciprocal chromosomal translocations, leads to different forms of leukaemia in humans.
-
The crystal structure of regulated Abl closely resembles that seen in structures of regulated Src-family kinases, but with unique differences: binding of myristate to a hydrophobic pocket in the kinase domain induces a defined conformational change that enables intramolecular docking of the SH2 domain to the kinase domain, thereby enforcing an autoinhibited conformation.
-
Phosphorylation sites have recently been mapped in Bcr–Abl by mass spectrometry. Taking the crystal structure and functional data into consideration, these new results have implications for Abl regulation.
-
The amino-terminal autoinhibitory 'Cap' region did not show interpretable electron density in the crystal structure, but a detailed functional analysis identified a region that is evolutionarily conserved and essential for autoinhibition and a second region that is not conserved and is dispensable.
-
An unsuspected link between mutations that confer resistance to the small-molecule inhibitor STI-571 and enzyme activation indicates that Abl-like regulatory constraints are also operational in the oncoprotein Bcr–Abl.
Abstract
The prototypic non-receptor tyrosine kinase c-Abl is implicated in various cellular processes. Its oncogenic counterpart, the Bcr–Abl fusion protein, causes certain human leukaemias. Recent insights into the structure and regulation of the c-Abl and Bcr–Abl tyrosine kinases have changed the way we look at these enzymes.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$189.00 per year
only $15.75 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Hanks, S. K. Genomic analysis of the eukaryotic protein kinase superfamily: a perspective. Genome Biol. 4, 111 (2003).
Pendergast, A. M. The Abl family kinases: mechanisms of regulation and signaling. Adv. Cancer Res. 85, 51–100 (2002). An extremely comprehensive review article covering, in particular, all aspects of Abl signalling mechanisms and cellular functions.
Woodring, P. J., Hunter, T. & Wang, J. Y. Regulation of F-actin-dependent processes by the Abl family of tyrosine kinases. J. Cell Sci. 116, 2613–2626 (2003).
Smith, J. M. & Mayer, B. J. Abl: mechanisms of regulation and activation. Front. Biosci. 7, d31–d42 (2002).
Advani, A. S. & Pendergast, A. M. Bcr–Abl variants: biological and clinical aspects. Leuk. Res. 26, 713–720 (2002).
Druker, B. J. et al. Chronic myelogenous leukemia. Hematology (Am. Soc. Hematol. Educ. Program) 87–112 (2001).
Van Etten, R. A. Cycling, stressed-out and nervous: cellular functions of c-Abl. Trends Cell Biol. 9, 179–186 (1999).
Abelson, H. T. & Rabstein, L. S. Lymphosarcoma: virus-induced thymic-independent disease in mice. Cancer Res. 30, 2213–2222 (1970). The historic paper describing the identification of v-Abl from Moloney murine leukaemia virus.
Abelson, H. T. & Rabstein, L. S. Influence of prednisolone on Moloney leukemogenic virus in BALB-c mice. Cancer Res. 30, 2208–2212 (1970).
Wang, J. Y. & Baltimore, D. Cellular RNA homologous to the Abelson murine leukemia virus transforming gene: expression and relationship to the viral sequence. Mol. Cell. Biol. 3, 773–779 (1983).
Raitano, A. B., Whang, Y. E. & Sawyers, C. L. Signal transduction by wild-type and leukemogenic Abl proteins. Biochim. Biophys. Acta 1333, F201–F216 (1997).
Daley, G. Q., Van Etten, R. A. & Baltimore, D. Induction of chronic myelogenous leukemia in mice by the P210bcr/abl gene of the Philadelphia chromosome. Science 247, 824–830 (1990).
Goldman, J. M. & Melo, J. V. Chronic myeloid leukemia — advances in biology and new approaches to treatment. N. Engl. J. Med. 349, 1451–1464 (2003).
McWhirter, J. R., Galasso, D. L. & Wang, J. Y. A coiled-coil oligomerization domain of Bcr is essential for the transforming function of Bcr–Abl oncoproteins. Mol. Cell. Biol. 13, 7587–7595 (1993).
Zhang, X., Subrahmanyam, R., Wong, R., Gross, A. W. & Ren, R. The NH(2)-terminal coiled-coil domain and tyrosine 177 play important roles in induction of a myeloproliferative disease in mice by Bcr–Abl. Mol. Cell. Biol. 21, 840–853 (2001).
Zhao, X., Ghaffari, S., Lodish, H., Malashkevich, V. N. & Kim, P. S. Structure of the Bcr–Abl oncoprotein oligomerization domain. Nature Struct. Biol. 7, 117–120 (2002).
Smith, K. M. & Van Etten, R. A. Activation of c-Abl kinase activity and transformation by a chemical inducer of dimerization. J. Biol. Chem. 276, 24372–24379 (2001).
Superti-Furga, G. & Courtneidge, S. A. Structure-function relationships in Src family and related protein tyrosine kinases. Bioessays 17, 321–330 (1995).
Smith, C. I. et al. The Tec family of cytoplasmic tyrosine kinases: mammalian Btk, Bmx, Itk, Tec, Txk and homologs in other species. Bioessays 23, 436–446 (2001).
Feller, S. M., Knudsen, B. & Hanafusa, H. c-Abl kinase regulates the protein binding activity of c-Crk. EMBO J. 13, 2341–2351 (1994).
Smith, J. M., Katz, S. & Mayer, B. J. Activation of the abl tyrosine kinase in vivo by src homology 3 domains from the src homology 2/Src homology 3 adaptor Nck. J. Biol. Chem. 274, 27956–27962 (1999).
Shafman, T. et al. Interaction between ATM protein and c-Abl in response to DNA damage. Nature 387, 520–523 (1997).
Goga, A., et al. p53 dependent growth suppression by the c-Abl nuclear tyrosine kinase. Oncogene 11, 791–799 (1995).
Welch, P. J. & Wang, J. Y. J. A c-terminal protein-binding domain in the retinoblastoma protein regulates nuclear c-abl tyrosine kinase in the cell-cycle. Cell 75, 779–790 (1993).
Baskaran, R., Chiang, G. G. & Wang, J. Y. Identification of a binding site in c-Abl tyrosine kinase for the C-terminal repeated domain of RNA polymerase II. Mol. Cell. Biol. 16, 3361–3369 (1996).
David-Cordonnier, M. H. et al. The DNA-binding domain of human c-Abl tyrosine kinase promotes the interaction of a HMG chromosomal protein with DNA. Nucleic Acids Res. 27, 2265–2270 (1999).
Van Etten, R. A. et al. The COOH terminus of the c-Abl tyrosine kinase contains distinct F- and G-actin binding domains with bundling activity. J. Cell Biol. 124, 325–340 (1994).
Van Etten, R. A., Jackson, P. & Baltimore, D. The mouse type IV c-abl gene product is a nuclear protein, and activation of transforming ability is associated with cytoplasmic localization. Cell 58, 669–678 (1989).
Taagepera, S. et al. Nuclear-cytoplasmic shuttling of C-ABL tyrosine kinase. Proc. Natl Acad. Sci. USA 95, 7457–7462 (1998).
Goga, A. et al. Oncogenic activation of c-ABL by mutation within its last exon. Mol. Cell. Biol. 13, 4967–4975 (1993).
Pluk, H., Dorey, K. & Superti-Furga, G. Autoinhibition of c-Abl. Cell 108, 247–259 (2002).
Nagar, B. et al. Structural basis for the autoinhibition of c-Abl tyrosine kinase. Cell 112, 859–871 (2003).
Harrison, S. C. Variation on an Src-like theme. Cell 112, 737–740 (2003).
Courtneidge, S. A. Cancer: escape from inhibition. Nature 422, 827–828 (2003).
Xu, W., Harrison, S. C. & Eck, M. J. Three-dimensional structure of the tyrosine kinase c-Src. Nature 385, 595–601 (1997).
Sicheri, F., Moarefi, I. & Kuriyan, J. Crystal structure of the Src family tyrosine kinase Hck. Nature 385, 602–609 (1997).
Williams, J. C. et al. The 2.35 Å crystal structure of the inactivated form of chicken Src: a dynamic molecule with multiple regulatory interactions. J. Mol. Biol. 274, 757–775 (1997).
Barilá, D. & Superti-Furga, G. An intramolecular SH3-domain interaction regulates c-Abl activity. Nature Genet. 18, 280–282 (1998). The article that first suggested c-Abl to be regulated by an intramolecular interaction involving the SH3 domain, the SH2–kinase linker and the small lobe. The structural model proposed, which includes a unique salt-bridge between the SH3 domain and the small lobe, is basically identical to the structure of regulated c-Abl determined five years later.
Young, M. A., Gonfloni, S., Superti-Furga, G., Roux, B. & Kuriyan, J. Dynamic coupling between the SH2 and SH3 domains of c-Src and Hck underlies their inactivation by C-terminal tyrosine phosphorylation. Cell 105, 115–126 (2001).
Hantschel, O. et al. A myristoyl/phosphotyrosine switch regulates c-Abl. Cell 112, 845–857 (2003). References 32 and 40 show the structure of regulated c-Abl, its autoinhibition mechanism, binding of the myristoyl group to the kinase domain, SH2 domain occlusion, activation by phosphotyrosine and conformational dependence for STI-571 binding.
Barilá, D. et al. A nuclear tyrosine phosphorylation circuit: c-Jun as an activator and substrate of c-Abl and JNK. EMBO J. 19, 273–281 (2000).
Juang, J. L. & Hoffmann, F. M. Drosophila Abelson interacting protein (dAbi) is a positive regulator of Abelson tyrosine kinase activity. Oncogene 18, 5138–5147 (1999).
Lewis, J. M. & Schwartz, M. A. Integrins regulate the association and phosphorylation of paxillin by c-Abl. J. Biol. Chem. 273, 14225–14230 (1998).
Miyoshi-Akiyama, T., Aleman, L. M., Smith, J. M., Adler, C. E. & Mayer, B. J. Regulation of Cbl phosphorylation by the Abl tyrosine kinase and the Nck SH2/SH3 adaptor. Oncogene 20, 4058–4069 (2001).
Roig, J., Tuazon, P. T., Zipfel, P. A., Pendergast, A. M. & Traugh, J. A. Functional interaction between c-Abl and the p21-activated protein kinase γ-PAK. Proc. Natl Acad. Sci. USA 97, 14346–14351 (2000).
Zukerberg, L. R. et al. Cables links Cdk5 and c-Abl and facilitates Cdk5 tyrosine phosphorylation, kinase upregulation, and neurite outgrowth. Neuron 26, 633–646 (2000).
Schindler, T. et al. Structural mechanism for STI-571 inhibition of abelson tyrosine kinase. Science 289, 1938–1942 (2000). Showed the structural basis for STI-571 inhibition and the first structure of the Abl kinase domain.
Nagar, B. et al. Crystal structures of the kinase domain of c-Abl in complex with the small molecule inhibitors PD173955 and imatinib (STI-571). Cancer Res. 62, 4236–4243 (2002).
Huse, M. & Kuriyan, J. The conformational plasticity of protein kinases. Cell 109, 275–282 (2002). A hallmark review for readers interested in the structural basis for the diverse mechanisms of protein kinase inhibition.
Yamaguchi, H. & Hendrickson, W. A. Structural basis for activation of human lymphocyte kinase Lck upon tyrosine phosphorylation. Nature 384, 484–489 (1996).
Franz, W. M., Berger, P. & Wang, J. Y. J. Deletion of an N-terminal regulatory domain of the c-Abl tyrosine kinase activates its oncogenic potential. EMBO J. 8, 137–147 (1989).
Daley, G. Q., Van Etten, R. A., Jackson, P. K., Bernards, A. & Baltimore, D. Nonmyristoylated Abl proteins transform a factor-dependent hematopoietic cell line. Mol. Cell. Biol. 12, 1864–1871 (1992).
Brasher, B. B. & Van Etten, R. A. c-Abl has high intrinsic tyrosine kinase activity that is stimulated by mutation of the src homology 3 domain and by autophosphorylation at two distinct regulatory tyrosines. J. Biol. Chem. 275, 35631–35637 (2000).
Dorey, K. et al. Phosphorylation and structure-based functional studies reveal a positive and a negative role for the activation loop of the c-Abl tyrosine kinase. Oncogene 20, 8075–8084 (2001).
Jackson, P. & Baltimore, D. N-terminal mutations activate the leukemogenic potential of the myristoylated form of c-Abl. EMBO J. 8, 449–456 (1989).
Van Etten, R. A., Debnath, J., Zhou, H. & Casasnovas, J. M. Introduction of a loss-of-function point mutation from the SH3 region of the Caenorhabditis elegans Sem-5 gene activates the transforming ability of c-Abl in vivo and abolishes binding of proline-rich ligands in vitro. Oncogene 10, 1977–1988 (1995).
Reynolds, F. H. Jr., Oroszlan, S. & Stephenson, J. R. Abelson murine leukemia virus P120: identification and characterization of tyrosine phosphorylation sites. J. Virol. 44, 1097–1101 (1982).
Plattner, R., Kadlec, L., DeMali, K. A., Kazlauskas, A. & Pendergast, A. M. c-Abl is activated by growth factors and src family kinases and has a role in the cellular response to PDGF. Genes Dev. 13, 2400–2411 (1999).
Furstoss, O. et al. c-Abl is an effector of Src for growth factor-induced c-myc expression and DNA synthesis. EMBO J. 21, 514–524 (2002).
Tanis, K. Q., Veach, D., Duewel, H. S., Bornmann, W. G. & Koleske, A. J. Two distinct phosphorylation pathways have additive effects on Abl family kinase activation. Mol. Cell. Biol. 23, 3884–3896 (2003).
Steen, H., Fernandez, M., Ghaffari, S., Pandey, A. & Mann, M. Phosphotyrosine mapping in Bcr/Abl oncoprotein using phosphotyrosine-specific immonium ion scanning. Mol. Cell Proteomics 2, 138–145 (2003).
Salomon, A. R. et al. Profiling of tyrosine phosphorylation pathways in human cells using mass spectrometry. Proc. Natl Acad. Sci. USA 100, 443–448 (2003).
Pendergast, A. M. et al. BCR–ABL-induced oncogenesis is mediated by direct interaction with the SH2 domain of the GRB-2 adaptor protein. Cell 75, 175–185 (1993).
Gould, K. L. & Nurse, P. Tyrosine phosphorylation of the fission yeast cdc2+ protein kinase regulates entry into mitosis. Nature 342, 39–45 (1989).
McGowan, C. H. & Russell, P. Human Wee1 kinase inhibits cell division by phosphorylating p34cdc2 exclusively on Tyr15. EMBO J. 12, 75–85 (1993).
Roumiantsev, S. et al. Clinical resistance to the kinase inhibitor STI-571 in chronic myeloid leukemia by mutation of Tyr-253 in the Abl kinase domain P-loop. Proc. Natl Acad. Sci. USA 99, 10700–10705 (2002). Contains the first evidence for a correlation between regulatory mutations and drug resistance.
Azam, M., Latek, R. R. & Daley, G. Q. Mechanisms of autoinhibition and STI-571/Imatinib resistance revealed by mutagenesis of BCR–ABL. Cell 112, 831–843 (2003). This groundbreaking paper changed the view of how Bcr–Abl functions, and will guide Abl researchers for the following years.
Allen, P. B. & Wiedemann, L. M. An activating mutation in the ATP binding site of the ABL kinase domain. J. Biol. Chem. 271, 19585–19591 (1996).
Brasher, B. B., Roumiantsev, S. & Van Etten, R. A. Mutational analysis of the regulatory function of the c-Abl Src homology 3 domain. Oncogene 20, 7744–7752 (2001).
Musacchio, A., Wilmanns, M. & Saraste, M. Structure and function of the SH3 domain. Prog. Biophys. Mol. Biol. 61, 283–297 (1994).
Zhou, H. X. How often does the myristoylated N-terminal latch of c-Abl come off? FEBS Lett. 552, 160–162 (2003).
Wright, P. E. & Dyson, H. J. Intrinsically unstructured proteins: re-assessing the protein structure-function paradigm. J. Mol. Biol. 293, 321–331 (1999).
Smith, K. M., Yacobi, R. & Van Etten, R. A. Autoinhibition of Bcr–Abl through its SH3 domain. Mol. Cell 12, 27–37 (2003).
Plattner, R. et al. A new link between the c-Abl tyrosine kinase and phosphoinositide signalling through PLC-γ1. Nature Cell Biol. 5, 309–319 (2003).
Plattner, R. & Pendergast, A. M. Activation and signaling of the Abl tyrosine kinase: bidirectional link with phosphoinositide signaling. Cell Cycle 2, 273–274 (2003).
Van Etten, R. A. c-Abl regulation: a tail of two lipids. Curr. Biol. 13, R608–R610 (2003).
Woodring, P. J., Hunter, T. & Wang, J. Y. Inhibition of c-Abl tyrosine kinase activity by filamentous actin. J. Biol. Chem. 276, 27104–27110 (2001).
Woodring, P. J. et al. Modulation of the F-actin cytoskeleton by c-Abl tyrosine kinase in cell spreading and neurite extension. J. Cell Biol. 156, 879–892 (2002).
Wen, S. -T. & Van Etten, R. A. The PAG gene product, a stress-induced protein with antioxidant properties, is an Abl SH3-binding protein and a physiological inhibitor of c-Abl tyrosine kinase activity. Genes Dev. 11, 2456–2467 (1997).
Neumann, C. A. et al. Essential role for the peroxiredoxin Prdx1 in erythrocyte antioxidant defence and tumour suppression. Nature 424, 561–565 (2003).
Baskaran, R. et al. Ataxia telangiectasia mutant protein activates c-Abl tyrosine kinase in response to ionizing radiation. Nature 387, 516–519 (1997).
Kharbanda, S. et al. Functional interaction between DNA-PK and c-Abl in response to DNA damage. Nature 386, 732–735 (1997).
Bhatnagar, R. S. et al. Structure of N-myristoyltransferase with bound myristoylCoA and peptide substrate analogs. Nature Struct. Biol. 5, 1091–1097 (1998).
Farazi, T. A., Waksman, G. & Gordon, J. I. The biology and enzymology of protein N–myristoylation. J. Biol. Chem. 276, 39501–39504 (2001).
Resh, M. D. Fatty acylation of proteins: new insights into membrane targeting of myristoylated and palmitoylated proteins. Biochim. Biophys. Acta 1451, 1–16 (1999). An extremely interesting review discussing the different mechanisms and functions of protein myristoylation and palmitoylation.
Resh, M. D. Myristylation and palmitylation of Src family members: the fats of the matter. Cell 76, 411–413 (1994).
Nicholson, D. W. Caspase structure, proteolytic substrates, and function during apoptotic cell death. Cell Death Differ. 6, 1028–1042 (1999).
Manning, G., Whyte, D. B., Martinez, R., Hunter, T. & Sudarsanam, S. The protein kinase complement of the human genome. Science 298, 1912–1934 (2002).
Cox, S., Radzio-Andzelm, E. & Taylor, S. S. Domain movements in protein kinases. Curr. Opin. Struct. Biol. 4, 893–901 (1994).
Buchdunger, E. et al. Inhibition of the Abl protein-tyrosine kinase in vitro and in vivo by a 2-phenylaminopyrimidine derivative. Cancer Res. 56, 100–104 (1996). First report about the selective inhibition of Abl and PDGF-R by STI-571.
Druker, B. J. et al. Effects of a selective inhibitor of the Abl tyrosine kinase on the growth of Bcr–Abl positive cells. Nature Med. 2, 561–566 (1996).
Druker, B. J. et al. Efficacy and safety of a specific inhibitor of the BCR–ABL tyrosine kinase in chronic myeloid leukemia. N. Engl. J. Med. 344, 1031–1037 (2001).
Heinrich, M. C. et al. Inhibition of c-kit receptor tyrosine kinase activity by STI-571, a selective tyrosine kinase inhibitor. Blood 96, 925–932 (2000).
Heinrich, M. C. et al. PDGFRA activating mutations in gastrointestinal stromal tumors. Science 299, 708–710 (2003).
Hochhaus, A. et al. Molecular and chromosomal mechanisms of resistance to imatinib (STI571) therapy. Leukemia 16, 2190–2196 (2002).
Mayer, B. J. SH3 domains: complexity in moderation. J. Cell Sci. 114, 1253–1263 (2001).
Pawson, T., Gish, G. D. & Nash, P. SH2 domains, interaction modules and cellular wiring. Trends Cell Biol. 11, 504–511 (2001).
Lemmon, M. A. Phosphoinositide recognition domains. Traffic 4, 201–213 (2003).
DeLano, W. L. The PyMOL Molecular Graphics System. The PyMOL homepage. <http://pymol.sourceforge.net.> (2002).
Acknowledgements
We are particularly grateful to K. Dorey and G. Neubauer for the unpublished data on Abl phosphorylation sites and K. Dorey for the sequence of Xenopus Abl. We would like to thank B. Nagar, K. Dorey, M. Schwab, A. Nebreda and K. Scheffzek for critical reading of the manuscript and J. Kuriyan and members of the Superti-Furga laboratory for insightful discussions. O.H. is supported by fellowships from the EMBL and the Aventis Foundation and is a fellow of the German National Merit Foundation. Work in the G.S.F. laboratory is supported by the EMBL and Cellzome.
Author information
Authors and Affiliations
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Glossary
- PARALOGUE
-
A gene product on the opposite branch of a duplicated gene family. Orthologues are on the same branch of a family (for example, FLNa, FLNb and FLNc). Paralogues and orthologues are homologues.
- PALMITOYLATION
-
Protein palmitoylation is a common protein modification in which a 16-carbon-atom saturated fatty acid (palmitate) is covalently attached to a cysteine residue through a thioester bond.
- SH2 DOMAIN
-
(Src-homology-2 domain). A protein module that recognizes and binds tyrosine-phosphorylated sequences in a sequence-specific context and thereby has a key role in relaying cascades of signal transduction.
- SH3 DOMAIN
-
(Src-homology-3 domain). A protein module of about 50 amino acids that recognizes and binds to sequences typically rich in proline.
- TEC-HOMOLOGY DOMAIN
-
A characteristic protein module found on Tec-family kinases, consisting of a highly conserved Zn2+ -binding Btk motif and a proline-rich region.
- PLECKSTRIN HOMOLOGY DOMAIN
-
A module of 100 amino acids that is present in many signalling molecules and binds to lipid products of phosphatidylinositol 3-kinase.
- ADAPTOR PROTEINS
-
Proteins that modulate cellular responses by recruiting other proteins to a complex. They usually contain several protein–protein interaction domains.
- ACTIVE SITE
-
The part of an enzyme where the substrates are brought into close proximity and the chemical reaction happens.
- POLYPROLINE TYPE II HELIX
-
A preferred conformation for proline-rich regions of protein sequences, with an axial translation of 3.2 Å and three residues in each turn of a left-handed helix. Other common polypeptide conformations are α-helix and β-sheet.
- ACTIVATION LOOP
-
A conserved structural motif in kinase domains, which needs to be phosphorylated for full activation of the kinase.
- SF9 CELLS
-
A commonly used insect cell line, derived from the fall armyworm (Spodoptera frugiperda), and used for baculovirus-mediated expression of recombinant proteins in insect cells.
- RT-LOOP
-
A variable loop in SH3 domains that is positioned close to conserved residues and is implicated in the binding of proline-rich (PXXP) motifs. It is responsible for differential binding affinity and specificity of SH3 domains.
- 310 HELIX
-
A tighter, less stable helix than the α-helix, with three residues per turn, forming hydrogen-bonded loops of 10 atoms.
- YEAST TWO-HYBRID
-
A technique used to test if two proteins physically interact with each other. One protein is fused to the GAL4 activation domain and the other to the GAL4 DNA-binding domain, and both fusion proteins are introduced into yeast. Expression of a GAL4-regulated reporter gene indicates that the two proteins physically interact.
Rights and permissions
About this article
Cite this article
Hantschel, O., Superti-Furga, G. Regulation of the c-Abl and Bcr–Abl tyrosine kinases. Nat Rev Mol Cell Biol 5, 33–44 (2004). https://doi.org/10.1038/nrm1280
Issue Date:
DOI: https://doi.org/10.1038/nrm1280