WW domain-containing proteins, WWOX and YAP, compete for interaction with ErbB-4 and modulate its transcriptional function.

The WW domain-containing oxidoreductase, WWOX, is a tumor suppressor that is deleted or altered in several cancer types. We recently showed that WWOX interacts with p73 and AP-2gamma and suppresses their transcriptional activity. Yes-associated protein (YAP), also containing WW domains, was shown to associate with p73 and enhance its transcriptional activity. In addition, YAP interacts with ErbB-4 receptor tyrosine kinase and acts as transcriptional coactivator of the COOH-terminal fragment (CTF) of ErbB-4. Stimulation of ErbB-4-expressing cells with 12-O-tetradecanoylphorbol-13-acetate (TPA) results in the proteolytic cleavage of its cytoplasmic domain and translocation of this domain to the nucleus. Here we report that WWOX physically associates with the full-length ErbB-4 via its first WW domain. Coexpression of WWOX and ErbB-4 in HeLa cells followed by treatment with TPA results in the retention of ErbB-4 in the cytoplasm. Moreover, in MCF-7 breast carcinoma cells, expressing high levels of endogenous WWOX, endogenous ErbB-4 is also retained in the cytoplasm. In addition, our results show that interaction of WWOX and ErbB-4 suppresses transcriptional coactivation of CTF by YAP in a dose-dependent manner. A mutant form of WWOX lacking interaction with ErbB-4 has no effect on this coactivation of ErbB-4. Furthermore, WWOX is able to inhibit coactivation of p73 by YAP. In summary, our data indicate that WWOX antagonizes the function of YAP by competing for interaction with ErbB-4 and other targets and thus affect its transcriptional activity.

[1]  Dimitrios Iliopoulos,et al.  Fragile genes as biomarkers: epigenetic control of WWOX and FHIT in lung, breast and bladder cancer , 2005, Oncogene.

[2]  J. Isola,et al.  Cleavable ErbB4 isoform in estrogen receptor-regulated growth of breast cancer cells. , 2005, Cancer research.

[3]  R. Weigel,et al.  Physical and Functional Interactions between the Wwox Tumor Suppressor Protein and the AP-2γ Transcription Factor , 2004, Cancer Research.

[4]  J. Ludes-Meyers,et al.  WWOX binds the specific proline-rich ligand PPXY: identification of candidate interacting proteins , 2004, Oncogene.

[5]  R. Roskoski.  The ErbB/HER receptor protein-tyrosine kinases and cancer. , 2004, Biochemical and biophysical research communications.

[6]  R. Roskoski.  The ErbB/HER receptor protein-tyrosine kinases and cancer*1 , 2004 .

[7]  Y. Pekarsky,et al.  Loss of WWOX Expression in Gastric Carcinoma , 2004, Clinical Cancer Research.

[8]  D. Iliopoulos,et al.  The fragile genes FHIT and WWOX are inactivated coordinately in invasive breast carcinoma , 2004, Cancer.

[9]  C. Croce,et al.  The Tumor Suppressor Gene WWOX at FRA16D Is Involved in Pancreatic Carcinogenesis , 2004, Clinical Cancer Research.

[10]  L. Frati,et al.  Ligand-regulated association of ErbB-4 to the transcriptional co-activator YAP65 controls transcription at the nuclear level. , 2004, Experimental cell research.

[11]  Y. Pekarsky,et al.  Functional association between Wwox tumor suppressor protein and p73, a p53 homolog. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[12]  Yi Zhang,et al.  A map of WW domain family interactions , 2004, Proteomics.

[13]  S. K. Zaidi,et al.  Tyrosine phosphorylation controls Runx2‐mediated subnuclear targeting of YAP to repress transcription , 2004, The EMBO journal.

[14]  J. Heath,et al.  Molecular mechanisms underlying WOX1 activation during apoptotic and stress responses. , 2003, Biochemical pharmacology.

[15]  N. Navin,et al.  WW Domain-containing Protein YAP Associates with ErbB-4 and Acts as a Co-transcriptional Activator for the Carboxyl-terminal Fragment of ErbB-4 That Translocates to the Nucleus* , 2003, Journal of Biological Chemistry.

[16]  G. Carpenter.  Nuclear localization and possible functions of receptor tyrosine kinases. , 2003, Current opinion in cell biology.

[17]  G. Carpenter,et al.  ErbB-4: mechanism of action and biology. , 2003, Experimental cell research.

[18]  B. Henderson,et al.  Regulation of tumor suppressors by nuclear-cytoplasmic shuttling. , 2003, Experimental cell research.

[19]  M. Fortini,et al.  Signalling: γ-Secretase-mediated proteolysis in cell-surface-receptor signalling , 2002, Nature Reviews Molecular Cell Biology.

[20]  J. Turkson,et al.  Cytoplasmic transport of Stat3 by receptor‐mediated endocytosis , 2002, The EMBO journal.

[21]  C. Croce,et al.  Genetic alterations of the tumor suppressor gene WWOX in esophageal squamous cell carcinoma. , 2002, Cancer research.

[22]  Gianni Cesareni,et al.  Normalization of nomenclature for peptide motifs as ligands of modular protein domains , 2002, FEBS letters.

[23]  A. Brenner,et al.  WWOX, the FRA16D gene, behaves as a suppressor of tumor growth. , 2001, Cancer research.

[24]  T. Golde,et al.  γ-Secretase Cleavage and Nuclear Localization of ErbB-4 Receptor Tyrosine Kinase , 2001, Science.

[25]  K. Makino,et al.  Nuclear localization of EGF receptor and its potential new role as a transcription factor , 2001, Nature Cell Biology.

[26]  D J Porteous,et al.  WWOX: A candidate tumor suppressor gene involved in multiple tumor types , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[27]  M. DePamphilis,et al.  TEAD/TEF transcription factors utilize the activation domain of YAP65, a Src/Yes-associated protein localized in the cytoplasm. , 2001, Genes & development.

[28]  L. Castagnoli,et al.  Physical Interaction with Yes-associated Protein Enhances p73 Transcriptional Activity* , 2001, The Journal of Biological Chemistry.

[29]  Y. Yarden,et al.  Untangling the ErbB signalling network , 2001, Nature Reviews Molecular Cell Biology.

[30]  T. Hunter,et al.  NeW Wrinkles for an Old Domain , 2000, Cell.

[31]  Xin Huang,et al.  Structure of a WW domain containing fragment of dystrophin in complex with β-dystroglycan , 2000, Nature Structural Biology.

[32]  R. Richards,et al.  Common chromosomal fragile site FRA16D sequence: identification of the FOR gene spanning FRA16D and homozygous deletions and translocation breakpoints in cancer cells. , 2000, Human molecular genetics.

[33]  A. Bednarek,et al.  WWOX, a novel WW domain-containing protein mapping to human chromosome 16q23.3-24.1, a region frequently affected in breast cancer. , 2000, Cancer research.

[34]  M. Klagsbrun,et al.  A Natural ErbB4 Isoform That Does Not Activate Phosphoinositide 3-Kinase Mediates Proliferation but Not Survival or Chemotaxis* , 2000, The Journal of Biological Chemistry.

[35]  D. Barnes,et al.  Nuclear expression of the c-erbB-4/HER-4 growth factor receptor in invasive breast cancers. , 2000, Cancer research.

[36]  M. Sudol,et al.  Yes-Associated Protein 65 Localizes P62c-Yes to the Apical Compartment of Airway Epithelia by Association with Ebp50 , 1999, The Journal of cell biology.

[37]  Lin-Feng Chen,et al.  A WW domain‐containing Yes‐associated protein (YAP) is a novel transcriptional co‐activator , 1999, The EMBO journal.

[38]  M. Klagsbrun,et al.  Characterization of a naturally occurring ErbB4 isoform that does not bind or activate phosphatidyl inositol 3-kinase , 1999, Oncogene.

[39]  Marius Sudol,et al.  From Src Homology domains to other signaling modules: proposal of the `protein recognition code' , 1998, Oncogene.

[40]  M. Klagsbrun,et al.  A Novel Juxtamembrane Domain Isoform of HER4/ErbB4 , 1997, The Journal of Biological Chemistry.

[41]  M. Saraste,et al.  Structure of the WW domain of a kinase-associated protein complexed with a proline-rich peptide , 1996, Nature.

[42]  M. Sudol,et al.  The WW domain of Yes-associated protein binds a proline-rich ligand that differs from the consensus established for Src homology 3-binding modules. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[43]  P. Bork,et al.  Characterization of the Mammalian YAP (Yes-associated Protein) Gene and Its Role in Defining a Novel Protein Module, the WW Domain (*) , 1995, The Journal of Biological Chemistry.

[44]  P. Bork,et al.  The WW domain: a signalling site in dystrophin? , 1994, Trends in biochemical sciences.

[45]  M. Sudol,et al.  Yes-associated protein (YAP65) is a proline-rich phosphoprotein that binds to the SH3 domain of the Yes proto-oncogene product. , 1994, Oncogene.

[46]  R. Weigel,et al.  Physical and functional interactions between the Wwox tumor suppressor protein and the AP-2gamma transcription factor. , 2004, Cancer research.

[47]  A. Sahin,et al.  Frequent loss of WWOX expression in breast cancer: correlation with estrogen receptor status , 2004, Breast Cancer Research and Treatment.

[48]  J. Downward,et al.  Akt phosphorylates the Yes-associated protein, YAP, to induce interaction with 14-3-3 and attenuation of p73-mediated apoptosis. , 2003, Molecular cell.

[49]  C. Van Waes,et al.  Genes differentially expressed with malignant transformation and metastatic tumor progression of murine squamous cell carcinoma , 1997, Journal of cellular biochemistry. Supplement.

[50]  M. Sudol,et al.  Structure and function of the WW domain. , 1996, Progress in biophysics and molecular biology.