Abstract
The catalytic activity of the c-Abl tyrosine kinase is tightly regulated by its Src homology 3 (SH3) domain through a complex mechanism that may involve intramolecular binding to Pro242 in the linker region between the SH2 and catalytic domains as well as interactions with a trans-inhibitor. We analysed the effect of mutation or replacement of SH3 on c-Abl tyrosine kinase activity and transformation. Random mutagenesis of SH3 identified several novel point mutations that dysregulated c-Abl kinase activity in vivo, but the RT loop was insensitive to mutational activation. Activating SH3 mutations abolished binding of proline-rich SH3 ligands in vitro, while mutations at Ser140 in the connector between the SH3 and SH2 domains activated Abl kinase activity in vivo and in vitro but did not impair SH3 ligand-binding. Abl was regulated efficiently when its SH3 domain was replaced with a heterologous SH3 from c-Src that binds a different spectrum of proline-rich ligands, but not by substitution of a modular WW domain with similar ligand-binding specificity. These results suggest that the SH3 domain regulates Abl principally by binding to the atypical intramolecular ligand Pro242 rather than a canonical PxxP ligand. Coordination between the SH3 and SH2 domains mediated by the connector region may be required for regulation of Abl even in the absence of SH2 ligand binding.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Barila D, Superti-Furga G . 1998 Nat. Genet. 18: 280–282
Bedford MT, Chan DC, Leder P . 1997 EMBO J. 16: 2376–2383
Brasher BB, Van Etten RA . 2000 J. Biol. Chem. 275: 35631–35637
Cartwright CA, Eckhart W, Simon S, Kaplan PL . 1987 Cell 49: 83–91
Chan DC, Bedford MT, Leder P . 1996 EMBO J. 15: 1045–1054
Dai Z, Pendergast AM . 1995 Genes Dev. 9: 2569–2582
Daley GQ, Van Etten RA, Jackson PK, Bernards A, Baltimore D . 1992 Mol. Cell. Biol. 12: 1864–1871
Dalgarno DC, Botfield MC, Rickles RJ . 1997 Biopoly. 43: 383–400
Dorey K, Barila D, Gavin AC, Nebreda AR, Superti-Furga G . 1999 Biol. Chem. 380: 223–230
Erpel T, Superti-Furga G, Courtneidge SA . 1995 EMBO J. 14: 963–975
Feng S, Chen JK, Yu H, Simon JA, Schreiber SL . 1994 Science 266: 1241–1247
Feng S, Kasahara C, Rickles RJ, Schreiber SL . 1995 Proc. Natl. Acad. Sci. USA 92: 12408–12415
Finer MH, Dull TJ, Qin L, Farson D, Roberts M . 1994 Blood 83: 43–50
Franz WM, Berger P, Wang JYJ . 1989 EMBO J. 8: 137–147
Hirai H, Varmus HE . 1990 Mol. Cell. Biol. 10: 1307–1318
Hunter T . 1987 Cell 49: 1–4
Jackson P, Baltimore D . 1989 EMBO J. 8: 449–456
Kay BK, Williamson MP, Sudol M . 2000 FASEB J. 14: 231–241
Kmiecik TE, Shalloway D . 1987 Cell 49: 65–73
Lee JC, Hapel AJ, Ihle JN . 1982 J. Immunol. 128: 2393–2398
Leung DW, Chen E, Goeddel DV . 1989 Technique 1: 11–15
Li S, Ilaria RL, Million RP, Daley GQ, Van Etten RA . 1999 J. Exp. Med. 189: 1399–1412
Lim WA, Richards FM . 1994 Nature Struct. Biol. 1: 221–225
Liu X, Brodeur SR, Gish G, Songyang Z, Cantley LC, Laudano AP, Pawson T . 1993 Oncogene 8: 1119–1126
Mayer BJ, Baltimore D . 1994 Mol. Cell. Biol. 14: 2883–2894
Mayer BJ, Jackson PK, Van Etten RA, Baltimore D . 1992 Mol. Cell. Biol. 12: 609–618
Palacios R, Steinmetz M . 1985 Cell 41: 727–734
Pendergast AM, Muller AJ, Havlik MH, Clark R, McCormick F, Witte ON . 1991 Proc. Natl. Acad. Sci. USA 88: 5927–5931
Pisabarro MT, Serrano L . 1996 Biochemistry 35: 10634–10640
Piwinica-worms H, Saunders K, Roberts T, Smith A, Cheng S . 1987 Cell 49: 75–82
Potts WM, Reynolds AB, Lansing TJ, Parsons JT . 1988 Oncogene Res. 3: 343–355
Ren R, Mayer BJ, Cicchetti P, Baltimore D . 1993 Science 259: 1157–1161
Seidel-Dugan C, Meyer BE, Thomas SM, Brugge JS . 1992 Mol. Cell. Biol. 12: 1835–1845
Shi Y, Alin K, Goff SP . 1995 Genes Dev. 9: 2583–2597
Van Etten RA . 1999 Trends Cell Biol. 9: 179–186
Van Etten RA, Debnath J, Zhou H, Casasnovas JM . 1995 Oncogene 10: 1977–1988
Van Etten RA, Jackson P, Baltimore D . 1989 Cell 58: 669–678
Walkenhorst J, Goga A, Witte ON, Superti-Furga G . 1996 Oncogene 12: 1513–1520
Welch PJ, Wang JYJ . 1993 Cell 75: 779–790
Wen S-T, Van Etten RA . 1997 Genes Dev. 11: 2456–2467
Weng Z, Rickles RJ, Feng S, Richard S, Shaw AS, Schreiber SL, Brugge JS . 1995 Mol. Cell. Biol. 15: 5627–5634
Xu W, Harrison SC, Eck MJ . 1997 Nature 385: 595–602
Young MA, Gonfloni S, Superti-Furga G, Roux B, Kuriyan J . 2001 Cell 105: 115–126
Zhu J, Shore SK . 1996 Mol. Cell. Biol. 16: 7054–7062
Acknowledgements
The authors thank Dr Peter Jackson (Stanford University) for reading the manuscript and Dr Bruce Mayer (University of Connecticut) for reagents and helpful discussions. This work was supported in part by a Howard Hughes Predoctoral Fellowship to B Brasher and by NIH grant CA72465 to RA Van Etten. RA Van Etten is a Scholar of the Leukemia and Lymphoma Society of America and The Carl and Margaret Walter Scholar in Blood Research at Harvard Medical School.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Brasher, B., Roumiantsev, S. & Van Etten, R. Mutational analysis of the regulatory function of the c-Abl Src homology 3 domain. Oncogene 20, 7744–7752 (2001). https://doi.org/10.1038/sj.onc.1204978
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1204978
Keywords
This article is cited by
-
Elucidation of protein interactions necessary for the maintenance of the BCR–ABL signaling complex
Cellular and Molecular Life Sciences (2020)
-
Identification and characterization of activating ABL1 1b kinase mutations: impact on sensitivity to ATP-competitive and allosteric ABL1 inhibitors
Leukemia (2017)
-
Atomic view of the energy landscape in the allosteric regulation of Abl kinase
Nature Structural & Molecular Biology (2017)
-
The Hippo pathway kinase Lats2 prevents DNA damage-induced apoptosis through inhibition of the tyrosine kinase c-Abl
Cell Death & Differentiation (2013)
-
A unique set of SH3–SH3 interactions controls IB1 homodimerization
The EMBO Journal (2006)