Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

ErbB2 kinase domain is required for ErbB2 association with β-catenin

ErbB2의 kinase 영역이 β-catenin과 ErbB2의 결합에 필요하다

  • Ha, Nam-Chul (Research Institute for Drug Development, College of Pharmacy, Pusan National University) ;
  • Xu, Wanping (Urologic Oncology Branch, National Cancer Institute) ;
  • Neckers, Len (Urologic Oncology Branch, National Cancer Institute) ;
  • Jung, Yun-Jin (Research Institute for Drug Development, College of Pharmacy, Pusan National University)
  • 하남출 (부산대학교 약학대학 신약개발연구소) ;
  • 슈완핑 (미국 국립암연구소 비뇨종양학실) ;
  • 넥컬즈렌 (미국 국립암연구소 비뇨종양학실) ;
  • 정연진 (부산대학교 약학대학 신약개발연구소)
  • Published : 2007.03.30
Download PDF
⟨ Previous Next ⟩

Abstract

To investigate the region of ErbB2 for the $ErbB2-{\beta}-catenin$ interaction, a proteasome $resistant-{\beta}-catenin$ and various ErbB2 constructs were transfected in COS7 cells. ErbB2 proteins were immunoprecipitated, and coimmunoprecipitated ${\beta}-catenin$ was examined by Western blotting. ${\beta}-catenin$ coimmunoprecipitated with full length ErbB2. Of the truncated ErbB2 proteins DT (1-1123), DHC (1-1031) and DK (1-750), the ErbB2 constructs containing the kinase domain, DT and DHC, precipitated together with ${\beta}-catenin$ but DK containing no kinase domain did not. To further test the requirement of the kinase domain for ${\beta}-catenin-ErbB2$ interaction, the presence of ${\beta}-catenin$ in the immunocomplex was examined following transfection with an ErbB2 mutant (${\triangle}750-971$) whose kinase domain is internally deleted and subsequent immunoprecipitation of the ErbB2 mutant. ${\beta}-catenin$ was not detected in the immunocomplex. These results suggest that the ErbB2 kinase domain comprises a potential site for ${\beta}-catenin$ binding to the receptor tyrosine kinase.

${\beta}-catenin$과 결합하는 ErbB2의 부위를 조사하기 위하여 proteasome에 의하여 분해되지 않는 ${\beta}-catenin$과 다양한 ErbB2 construct를 COS7 세포에 transfection한 후 ErbB2 단백질을 그것의 항체로 가라앉혔다. 이 때 공침한 ${\beta}-catenin$을 Western blot으로 분석하였다. C 말단에서부터 잘려진 ErbB2 단백질 중에 kinase 영역을 가지고 있는 것들만 ${\beta}-catenin$과 공침하였다. kinase 영역의 필요성을 확인하기 위하여 kinase 영역이 내부에서 제거된 ErbB2 construct를 ${\beta}-catenin$과 transfection 한 후 동일한 실험을 실시하였다. 이 실험에서 ${\beta}-catenin$는 kinase 영역이 내부적으로 제거된 ErbB2 단백질과 공침하지 않았다. 이 결과는 ${\beta}-catenin$과 결합하는 ErbB2의 위치는 kinase 영역내에 있음을 제시한다.

Keywords

References

  1. Ozawa, M., H. Baribault and R. Kemler. 1989. The cytoplasmic domain of the cell adhesion molecule uvomorulin associates with three independent proteins structurally related in different species. EMBO J. 8, 1711-1717
  2. Peifer, M., P. D. McCrea, K. J. Green, E. Wieschaus and B. M. Gumbiner. 1992. The vertebrate adhesive junction proteins ${\beta}$-catenin and plakoglobin and the Drosophila segment polarity gene armadillo form a multigene family with similar properties. J. Cell. Biol. 118, 681-691 https://doi.org/10.1083/jcb.118.3.681
  3. Behrens, J., J. P. von Kries, M. Kuhl, L. Bruhn, D. Wedlich, R. Grosschedl and W. Birchmeier. 1996. Functional interaction of beta-catenin with the transcription factor LEF-1. Nature 382, 638-642 https://doi.org/10.1038/382638a0
  4. Molenaar, M., M. van de Wetering, M. Oosterwegel, J. Peterson-Maduro, S. Godsave, V. Korinek, J. Roose, O. Destree and H. Clevers. 1996. XTcf-3 transcription factor mediates ${\beta}$-catenin-induced axis formation in Xenopus embryos. Cell 86, 391-399 https://doi.org/10.1016/S0092-8674(00)80112-9
  5. Cavallo, R., D. Rubenstein and M. Peifer. 1997. Armadillo and dTCF: a marriage made in the nucleus. Curr. Opin. Genet. Dev. 7, 459-466 https://doi.org/10.1016/S0959-437X(97)80071-8
  6. Hatzfeld, M. 1999. The armadillo family of structural proteins. Int. Rev. Cytol. 186, 179-224
  7. Cowin, P., H. P. Kapprell, W. W. Franke, J. Tamkun and R. O. Hynes. 1986. Plakoglobin: a protein common to different kinds of intercellular adhering junctions. Cell 46, 1063-1073 https://doi.org/10.1016/0092-8674(86)90706-3
  8. McCrea, P. D., C. W. Turck and B. Gumbiner. 1991. A homolog of the armadillo protein in Drosophila (plakoglobin) associated with E-cadherin. Science 254, 1359-1361 https://doi.org/10.1126/science.1962194
  9. Kemler, R. 1993. From cadherins to catenins: cytoplasmic protein interactions and regulation of cell adhesion. Trends Genet. 9, 317-321 https://doi.org/10.1016/0168-9525(93)90250-L
  10. Drees, F., S. Pokutta, S. Yamada, W. J. Nelson and W. I. Weis. 2005. ${\alpha}$-catenin is a molecular switch that binds E-cadherin-${\beta}$-catenin and regulates actin-filament assembly. Cell 123, 903-915 https://doi.org/10.1016/j.cell.2005.09.021
  11. Yamada, S., S. Pokutta, F. Drees, W. I. Weis and W. J. Nelson. 2005. Deconstructing the cadherin-catenin-actin complex. Cell 123, 889-901 https://doi.org/10.1016/j.cell.2005.09.020
  12. Thiery, J. P. 2002. Epithelial-mesenchymal transitions in tumour progression. Nat. Rev. Cancer 2, 442-454 https://doi.org/10.1038/nrc822
  13. Behrens, J., M. M. Mareel, F. M. Van Roy and W. Birchmeier. 1989. Dissecting tumor cell invasion: epithelial cells acquire invasive properties after the loss of uvomorulin-mediated cell-cell adhesion. J. Cell Biol. 108, 2435-2447 https://doi.org/10.1083/jcb.108.6.2435
  14. Vleminckx, K., L. Vakaet, Jr., M. Mareel, W. Fiers and F. van Roy. 1991. Genetic manipulation of E-cadherin expression by epithelial tumor cells reveals an invasion suppressor role. Cell 66, 107-119 https://doi.org/10.1016/0092-8674(91)90143-M
  15. Tetsu, O. and F. McCormick. 1999. ${\beta}$-catenin regulates expression of cyclin D1 in colon carcinoma cells. Nature 398, 422-426 https://doi.org/10.1038/18884
  16. Polakis, P. 2000. Wnt signaling and cancer. Genes Dev. 14, 1837-1851
  17. Lilien, J. and J. Balsamo. 2005. The regulation of cadherin-mediated adhesion by tyrosine phosphorylation/dephosphorylation of ${\beta}$-catenin. Curr. Opin. Cell Biol. 17, 459-465 https://doi.org/10.1016/j.ceb.2005.08.009
  18. Wang, Z., D. Shen, D. W. Parsons, A. Bardelli, J. Sager, S. Szabo, J. Ptak, N. Silliman, B. A. Peters, M. S. van der Heijden, G. Parmigiani, H. Yan, T. L. Wang, G. Riggins, S. M. Powell, J. K. Willson, S. Markowitz, K. W. Kinzler, B. Vogelstein and V. E. Velculescu. 2004. Mutational analysis of the tyrosine phosphatome in colorectal cancers. Science 304, 1164-1166 https://doi.org/10.1126/science.1096096
  19. Hoschuetzky, H., H. Aberle and R. Kemler. 1994. ${\beta}$-catenin mediates the interaction of the cadherin-catenin complex with epidermal growth factor receptor. J. Cell Biol. 127, 1375-1380 https://doi.org/10.1083/jcb.127.5.1375
  20. Kanai, Y., A. Ochiai, T. Shibata, T. Oyama, S. Ushijima, S. Akimoto and S. Hirohashi. 1995. c-erbB-2 gene product directly associates with ${\beta}$-catenin and plakoglobin. Biochem. Biophys. Res. Commun. 208, 1067-1072 https://doi.org/10.1006/bbrc.1995.1443
  21. Shibata, T., A. Ochiai, Y. Kanai, S. Akimoto, M. Gotoh, N. Yasui, R. Machinami and S. Hirohashi. 1996. Dominant negative inhibition of the association between ${\beta}$-catenin and c-erbB-2 by N-terminally deleted ${\beta}$-catenin suppresses the invasion and metastasis of cancer cells. Oncogene 13, 883-889
  22. Roura, S., S. Miravet, J. Piedra, A. Garcia de Herreros and M. Dunach, 1999. Regulation of E-cadherin/Catenin association by tyrosine phosphorylation. J. Biol. Chem. 274, 36734-36740 https://doi.org/10.1074/jbc.274.51.36734
  23. Tzahar, E., H. Waterman, X. Chen, G. Levkowitz, D. Karunagaran, S. Lavi, B. J. Ratzkin and Y. Yarden. 1996. A hierarchical network of interreceptor interactions determines signal transduction by Neu differentiation factor/neuregulin and epidermal growth factor. Mol. Cell Biol. 16, 5276-5287 https://doi.org/10.1128/MCB.16.10.5276
  24. Chavany, C., E. Mimnaugh, P. Miller, R. Bitton, P. Nguyen, J. Trepel, L. Whitesell, R. Schnur, J. Moyer and L. Neckers. 1996. p185erbB2 binds to GRP94 in vivo. Dissociation of the p185erbB2/GRP94 heterocomplex by benzoquinone ansamycins precedes depletion of p185erbB2. J. Biol. Chem. 271, 4974-4977 https://doi.org/10.1074/jbc.271.9.4974
  25. Muller, T., A. Choidas, E. Reichmann and A. Ullrich, 1999. Phosphorylation and free pool of ${\beta}$-catenin are regulated by tyrosine kinases and tyrosine phosphatases during epithelial cell migration. J. Biol. Chem. 274, 10173-10183 https://doi.org/10.1074/jbc.274.15.10173
  26. Bonvini, P., W. G. An, A. Rosolen, P. Nguyen, J. Trepel, A. Garcia de Herreros, M. Dunach and L. M. Neckers. 2001. Geldanamycin abrogates ErbB2 association with proteasome-resistant ${\beta}$-catenin in melanoma cells, increases beta-catenin-E-cadherin association, and decreases ${\beta}$-catenin-sensitive transcription. Cancer Res. 61, 1671-1677
  27. Rubinfeld, B., P. Robbins, M. El-Gamil, I. Albert, E. Porfiri and P. Polakis. 1997. Stabilization of ${\beta}$-catenin by genetic defects in melanoma cell lines. Science 275, 1790-1792 https://doi.org/10.1126/science.275.5307.1790
  28. Sommers, C. L., E. P. Gelmann, R. Kemler, P. Cowin and S. W. Byers. 1994. Alterations in beta-catenin phosphorylation and plakoglobin expression in human breast cancer cells. Cancer Res. 54, 3544-3552