Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.


Logo for

J. Biol. Chem. 276 (37): 35185-35193

© 2001 by The American Society for Biochemistry and Molecular Biology, Inc.

Src-catalyzed Phosphorylation of c-Cbl Leads to the Interdependent Ubiquitination of Both Proteins*

Masahiro YokouchiDagger §||, Takeshi Kondo**, Archana SanjayDagger §Dagger Dagger , Adam HoughtonDagger §Dagger Dagger , Akihiko Yoshimura§§, Seturo Komiya, Hui Zhang**, and Roland BaronDagger §¶¶

From the Departments of Dagger  Cell Biology, § Orthopaedics, and ** Genetics, Yale University School of Medicine, New Haven, Connecticut 06511, the  Department of Orthopaedic Surgery, Faculty of Medicine, Kagoshima University, Kagoshima 890-8520, Japan, and §§ Institute of Life Science, Kurume University, Kurume 893-0861, Japan

The protooncogene c-Cbl has recently emerged as an E3 ubiquitin ligase for activated receptor tyrosine kinases. We report here that c-Cbl also mediates the ubiquitination of another protooncogene, the non-receptor tyrosine kinase c-Src, as well as of itself. The c-Cbl-dependent ubiquitination of Src and c-Cbl requires c-Cbl's RING finger, Src kinase activity, and c-Cbl's tyrosine phosphorylation, probably on Tyr-371. In vitro, c-Cbl forms a stable complex with the ubiquitin-conjugating enzyme UbcH7, but active Src destabilizes this interaction. In contrast, Src inhibition stabilizes the c-Cbl· UbcH7·Src complex. Finally, c-Cbl reduces v-Src protein levels and suppresses v-Src-induced STAT3 activation. Thus, in addition to mediating the ubiquitination of activated receptor tyrosine kinases, c-Cbl also acts as a ubiquitin ligase for the non-receptor tyrosine kinase Src, thereby down-regulating Src.

* This work was supported by Grant AR-42927 from the NIAMS and The Office of Research on Women's Health, National Institutes of Health (NIH) (to R.B.) and by NIH Grant CA-72878 (to H.Z.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

|| Recipient of fellowships from the Japan Society for the Promotion of Science and from the Hip Joint Foundation of Japan, Inc.

Dagger Dagger Recipients of post-doctoral fellowships from the Arthritis Foundation.

¶¶ To whom correspondence should be addressed: Depts. of Cell Biology and Orthopaedics, Yale University School of Medicine, 333 Cedar St., New Haven, CT 06510. Tel.: 203-785-4150; Fax: 203-785-2744; E-mail:

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.

A comprehensive pathway map of epidermal growth factor receptor signaling.
K. Oda, Y. Matsuoka, A. Funahashi, and H. Kitano (2014)
Mol Syst Biol 1, 2005.0010
   Abstract »    Full Text »    PDF »
Microtubule Dynamic Instability Controls Podosome Patterning in Osteoclasts through EB1, Cortactin, and Src.
M. Biosse Duplan, D. Zalli, S. Stephens, S. Zenger, L. Neff, J. M. Oelkers, F. P. L. Lai, W. Horne, K. Rottner, and R. Baron (2014)
Mol. Cell. Biol. 34, 16-29
   Abstract »    Full Text »    PDF »
Threshold-controlled ubiquitination of the EGFR directs receptor fate.
S. Sigismund, V. Algisi, G. Nappo, A. Conte, R. Pascolutti, A. Cuomo, T. Bonaldi, E. Argenzio, L. G. G. C. Verhoef, E. Maspero, et al. (2013)
EMBO J. 32, 2140-2157
   Abstract »    Full Text »    PDF »
Inhibition of SRC Corrects GM-CSF Hypersensitivity That Underlies Juvenile Myelomonocytic Leukemia.
S. Bunda, M. W. Kang, S. S. Sybingco, J. Weng, H. Favre, D. H. Shin, M. S. Irwin, M. L. Loh, and M. Ohh (2013)
Cancer Res. 73, 2540-2550
   Abstract »    Full Text »    PDF »
Src-dependent autophagic degradation of Ret in FAK-signalling-defective cancer cells.
E. Sandilands, B. Serrels, S. Wilkinson, and M. C. Frame (2012)
EMBO Rep. 13, 733-740
   Abstract »    Full Text »    PDF »
The ESCRT machinery mediates polarization of fibroblasts through regulation of myosin light chain.
V. H. Lobert and H. Stenmark (2012)
J. Cell Sci. 125, 29-36
   Abstract »    Full Text »    PDF »
Cell Polarity and Migration: Emerging Role for the Endosomal Sorting Machinery.
V. H. Lobert and H. Stenmark (2011)
Physiology 26, 171-180
   Abstract »    Full Text »    PDF »
The Src-Like Adaptor Protein Regulates GM-CSFR Signaling and Monocytic Dendritic Cell Maturation.
L. M. Liontos, D. Dissanayake, P. S. Ohashi, A. Weiss, L. L. Dragone, and C. J. McGlade (2011)
J. Immunol. 186, 1923-1933
   Abstract »    Full Text »    PDF »
LAB/NTAL/Lat2: a force to be reckoned with in all leukocytes?.
S. J. Orr and D. W. McVicar (2011)
J. Leukoc. Biol. 89, 11-19
   Abstract »    Full Text »    PDF »
Exendin-4 Suppresses Src Activation and Reactive Oxygen Species Production in Diabetic Goto-Kakizaki Rat Islets in an Epac-Dependent Manner.
E. Mukai, S. Fujimoto, H. Sato, C. Oneyama, R. Kominato, Y. Sato, M. Sasaki, Y. Nishi, M. Okada, and N. Inagaki (2011)
Diabetes 60, 218-226
   Abstract »    Full Text »    PDF »
As4S4 targets RING-type E3 ligase c-CBL to induce degradation of BCR-ABL in chronic myelogenous leukemia.
J.-H. Mao, X.-Y. Sun, J.-X. Liu, Q.-Y. Zhang, P. Liu, Q.-H. Huang, K. K. Li, Q. Chen, Z. Chen, and S.-J. Chen (2010)
PNAS 107, 21683-21688
   Abstract »    Full Text »    PDF »
Myristoylation and Membrane Binding Regulate c-Src Stability and Kinase Activity.
P. Patwardhan and M. D. Resh (2010)
Mol. Cell. Biol. 30, 4094-4107
   Abstract »    Full Text »    PDF »
The N Terminus of Cbl-c Regulates Ubiquitin Ligase Activity by Modulating Affinity for the Ubiquitin-conjugating Enzyme.
P. E. Ryan, N. Sivadasan-Nair, M. M. Nau, S. Nicholas, and S. Lipkowitz (2010)
J. Biol. Chem. 285, 23687-23698
   Abstract »    Full Text »    PDF »
Regulation of PTEN/Akt and MAP kinase signaling pathways by the ubiquitin ligase activators Ndfip1 and Ndfip2.
T. Mund and H. R. B. Pelham (2010)
PNAS 107, 11429-11434
   Abstract »    Full Text »    PDF »
Negative Feedback Control of Osteoclast Formation through Ubiquitin-mediated Down-regulation of NFATc1.
J. H. Kim, K. Kim, H. M. Jin, I. Song, B. U. Youn, S.-H. Lee, Y. Choi, and N. Kim (2010)
J. Biol. Chem. 285, 5224-5231
   Abstract »    Full Text »    PDF »
Down-regulation of c-Cbl by Morphine Accounts for Persistent ERK1/2 Signaling in {delta}-Opioid Receptor-expressing HEK293 Cells.
D. A. Eisinger and H. Ammer (2009)
J. Biol. Chem. 284, 34819-34828
   Abstract »    Full Text »    PDF »
E3 ligase-defective Cbl mutants lead to a generalized mastocytosis and myeloproliferative disease.
S. R. Bandi, C. Brandts, M. Rensinghoff, R. Grundler, L. Tickenbrock, G. Kohler, J. Duyster, W. E. Berdel, C. Muller-Tidow, H. Serve, et al. (2009)
Blood 114, 4197-4208
   Abstract »    Full Text »    PDF »
Breakdown of endocytosis in the oncogenic activation of receptor tyrosine kinases.
J. V. Abella and M. Park (2009)
Am J Physiol Endocrinol Metab 296, E973-E984
   Abstract »    Full Text »    PDF »
The phytochemical piceatannol induces the loss of CBL and CBL-associated proteins.
A. C. Klimowicz, S. A. Bisson, K. Hans, E. M. Long, H. C. Hansen, and S. M. Robbins (2009)
Mol. Cancer Ther. 8, 602-614
   Abstract »    Full Text »    PDF »
Herpes Simplex Virus Type 1 ICP0 Phosphorylation Mutants Impair the E3 Ubiquitin Ligase Activity of ICP0 in a Cell Type-Dependent Manner.
C. Boutell, R. Everett, J. Hilliard, P. Schaffer, A. Orr, and D. Davido (2008)
J. Virol. 82, 10647-10656
   Abstract »    Full Text »    PDF »
Suppression of v-Src Transformation by Andrographolide via Degradation of the v-Src Protein and Attenuation of the Erk Signaling Pathway.
F.-P. Liang, C.-H. Lin, C.-D. Kuo, H.-P. Chao, and S.-L. Fu (2008)
J. Biol. Chem. 283, 5023-5033
   Abstract »    Full Text »    PDF »
Srcasm Corrects Fyn-induced Epidermal Hyperplasia by Kinase Down-regulation.
W. Li, C. Marshall, L. Mei, J. Gelfand, and J. T. Seykora (2007)
J. Biol. Chem. 282, 1161-1169
   Abstract »    Full Text »    PDF »
KEEP ON GOING, a RING E3 Ligase Essential for Arabidopsis Growth and Development, Is Involved in Abscisic Acid Signaling.
S. L. Stone, L. A. Williams, L. M. Farmer, R. D. Vierstra, and J. Callis (2006)
PLANT CELL 18, 3415-3428
   Abstract »    Full Text »    PDF »
Dynamin Forms a Src Kinase-sensitive Complex with Cbl and Regulates Podosomes and Osteoclast Activity.
A. Bruzzaniti, L. Neff, A. Sanjay, W. C. Horne, P. De Camilli, and R. Baron (2005)
Mol. Biol. Cell 16, 3301-3313
   Abstract »    Full Text »    PDF »
Structural Characterization of a Novel Cbl Phosphotyrosine Recognition Motif in the APS Family of Adapter Proteins.
J. Hu and S. R. Hubbard (2005)
J. Biol. Chem. 280, 18943-18949
   Abstract »    Full Text »    PDF »
Cortactin Overexpression Inhibits Ligand-Induced Down-regulation of the Epidermal Growth Factor Receptor.
P. Timpson, D. K. Lynch, D. Schramek, F. Walker, and R. J. Daly (2005)
Cancer Res. 65, 3273-3280
   Abstract »    Full Text »    PDF »
Cbl-mediated ubiquitination of {alpha}5 integrin subunit mediates fibronectin-dependent osteoblast detachment and apoptosis induced by FGFR2 activation.
K. Kaabeche, H. Guenou, D. Bouvard, N. Didelot, A. Listrat, and P. J. Marie (2005)
J. Cell Sci. 118, 1223-1232
   Abstract »    Full Text »    PDF »
Regulation of stem cell factor receptor signaling by Cbl family proteins (Cbl-b/c-Cbl).
S. Zeng, Z. Xu, S. Lipkowitz, and J. B. Longley (2005)
Blood 105, 226-232
   Abstract »    Full Text »    PDF »
Apc Deficiency Is Associated with Increased Egfr Activity in the Intestinal Enterocytes and Adenomas of C57BL/6J-Min/+ Mice.
A. E. Moran, D. H. Hunt, S. H. Javid, M. Redston, A. M. Carothers, and M. M. Bertagnolli (2004)
J. Biol. Chem. 279, 43261-43272
   Abstract »    Full Text »    PDF »
Cbl-mediated Degradation of Lyn and Fyn Induced by Constitutive Fibroblast Growth Factor Receptor-2 Activation Supports Osteoblast Differentiation.
K. Kaabeche, J. Lemonnier, S. Le Mee, J. Caverzasio, and P. J. Marie (2004)
J. Biol. Chem. 279, 36259-36267
   Abstract »    Full Text »    PDF »
Biochemical Basis for the Requirement of Kinase Activity for Cbl-dependent Ubiquitinylation and Degradation of a Target Tyrosine Kinase.
A. K. Ghosh, A. L. Reddi, N. L. Rao, L. Duan, V. Band, and H. Band (2004)
J. Biol. Chem. 279, 36132-36141
   Abstract »    Full Text »    PDF »
Regulation of Ubiquitin Protein Ligase Activity in c-Cbl by Phosphorylation-induced Conformational Change and Constitutive Activation by Tyrosine to Glutamate Point Mutations.
C. K. Kassenbrock and S. M. Anderson (2004)
J. Biol. Chem. 279, 28017-28027
   Abstract »    Full Text »    PDF »
Regulation of the Src Family Kinase Lck by Hsp90 and Ubiquitination.
A. Giannini and M.-J. Bijlmakers (2004)
Mol. Cell. Biol. 24, 5667-5676
   Abstract »    Full Text »    PDF »
Ubiquitination and Proteasomal Degradation of Nucleophosmin-Anaplastic Lymphoma Kinase Induced by 17-Allylamino-Demethoxygeldanamycin: Role of the Co-Chaperone Carboxyl Heat Shock Protein 70-Interacting Protein.
P. Bonvini, H. D. Rosa, N. Vignes, and A. Rosolen (2004)
Cancer Res. 64, 3256-3264
   Abstract »    Full Text »    PDF »
Newest findings on the oldest oncogene; how activated src does it.
M. C. Frame (2004)
J. Cell Sci. 117, 989-998
   Abstract »    Full Text »    PDF »
WW Domain HECT E3s Target Cbl RING Finger E3s for Proteasomal Degradation.
A. Magnifico, S. Ettenberg, C. Yang, J. Mariano, S. Tiwari, S. Fang, S. Lipkowitz, and A. M. Weissman (2003)
J. Biol. Chem. 278, 43169-43177
   Abstract »    Full Text »    PDF »
Tyrosine Residues Direct the Ubiquitination and Degradation of the NY-1 Hantavirus G1 Cytoplasmic Tail.
E. Geimonen, I. Fernandez, I. N. Gavrilovskaya, and E. R. Mackow (2003)
J. Virol. 77, 10760-10868
   Abstract »    Full Text »    PDF »
High Precision NMR Structure and Function of the RING-H2 Finger Domain of EL5, a Rice Protein Whose Expression Is Increased upon Exposure to Pathogen-derived Oligosaccharides.
S. Katoh, C. Hong, Y. Tsunoda, K. Murata, R. Takai, E. Minami, T. Yamazaki, and E. Katoh (2003)
J. Biol. Chem. 278, 15341-15348
   Abstract »    Full Text »    PDF »
ARC1 Is an E3 Ubiquitin Ligase and Promotes the Ubiquitination of Proteins during the Rejection of Self-Incompatible Brassica Pollen.
S. L. Stone, E. M. Anderson, R. T. Mullen, and D. R. Goring (2003)
PLANT CELL 15, 885-898
   Abstract »    Full Text »    PDF »
A Novel RING Finger Protein, Human Enhancer of Invasion 10, Alters Mitotic Progression through Regulation of Cyclin B Levels.
G. G. Toby, W. Gherraby, T. R. Coleman, and E. A. Golemis (2003)
Mol. Cell. Biol. 23, 2109-2122
   Abstract »    Full Text »    PDF »
Src promotes destruction of c-Cbl: Implications for oncogenic synergy between Src and growth factor receptors.
J. Bao, G. Gur, and Y. Yarden (2003)
PNAS 100, 2438-2443
   Abstract »    Full Text »    PDF »
Ubiquitylation of MEKK1 Inhibits Its Phosphorylation of MKK1 and MKK4 and Activation of the ERK1/2 and JNK Pathways.
J. A. Witowsky and G. L. Johnson (2003)
J. Biol. Chem. 278, 1403-1406
   Abstract »    Full Text »    PDF »
Activation of Syk Tyrosine Kinase Is Required for c-Cbl-mediated Ubiquitination of Fc{varepsilon}RI and Syk in RBL Cells.
R. Paolini, R. Molfetta, L. O. Beitz, J. Zhang, A. M. Scharenberg, M. Piccoli, L. Frati, R. Siraganian, and A. Santoni (2002)
J. Biol. Chem. 277, 36940-36947
   Abstract »    Full Text »    PDF »
Sprouty2 attenuates epidermal growth factor receptor ubiquitylation and endocytosis, and consequently enhances Ras/ERK signalling.
E. S. M. Wong, C. W. Fong, J. Lim, P. Yusoff, B. C. Low, W. Y. Langdon, and G. R. Guy (2002)
EMBO J. 21, 4796-4808
   Abstract »    Full Text »    PDF »
Identification of Novel SH3 Domain Ligands for the Src Family Kinase Hck: WISKOTT-ALDRICH SYNDROME PROTEIN (WASP), WASP-INTERACTING PROTEIN (WIP), AND ELMO1.
M. P. Scott, F. Zappacosta, E. Y. Kim, R. S. Annan, and W. T. Miller (2002)
J. Biol. Chem. 277, 28238-28246
   Abstract »    Full Text »    PDF »
Inhibition of Src Family Kinases Blocks Epidermal Growth Factor (EGF)-induced Activation of Akt, Phosphorylation of c-Cbl, and Ubiquitination of the EGF Receptor.
C. K. Kassenbrock, S. Hunter, P. Garl, G. L. Johnson, and S. M. Anderson (2002)
J. Biol. Chem. 277, 24967-24975
   Abstract »    Full Text »    PDF »
Regulation of Jak2 through the Ubiquitin-Proteasome Pathway Involves Phosphorylation of Jak2 on Y1007 and Interaction with SOCS-1.
D. Ungureanu, P. Saharinen, I. Junttila, D. J. Hilton, and O. Silvennoinen (2002)
Mol. Cell. Biol. 22, 3316-3326
   Abstract »    Full Text »    PDF »
The Cbl family of ubiquitin ligases: critical negative regulators of tyrosine kinase signaling in the immune system.
N. Rao, I. Dodge, and H. Band (2002)
J. Leukoc. Biol. 71, 753-763
   Abstract »    Full Text »    PDF »
Negative regulation of Lck by Cbl ubiquitin ligase.
N. Rao, S. Miyake, A. L. Reddi, P. Douillard, A. K. Ghosh, I. L. Dodge, P. Zhou, N. D. Fernandes, and H. Band (2002)
PNAS 99, 3794-3799
   Abstract »    Full Text »    PDF »
The Cbl Family: Ubiquitin Ligases Regulating Signaling by Tyrosine Kinases.
A. Sanjay, W. C. Horne, and R. Baron (2001)
Sci. STKE 2001, pe40
   Abstract »    Full Text »    PDF »
The checkpoint protein Chfr is a ligase that ubiquitinates Plk1 and inhibits Cdc2 at the G2 to M transition.
D. Kang, J. Chen, J. Wong, and G. Fang (2002)
J. Cell Biol. 156, 249-260
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882