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Gene Fusion Proteins

Cytoplasmic and nuclear localization of the 130 and 160 kDa Bcr proteins

Leukemia volume 14, pages 1892–1897 (2000)Cite this article

Abstract

Formation of the Bcr-Abl chimeric protein is the molecular hallmark of Philadelphia-positive leukemia. Normal Bcr is a complex protein which has been found in the cytoplasm, has serine kinase activity, and has been implicated in cellular signal transduction. However, we have recently demonstrated that Bcr can also associate with condensed chromatin. Since two major Bcr proteins have been characterized (p160Bcr and p130Bcr), we sought to determine if different forms of Bcr localized to the nucleus vs the cytoplasm. Metabolic labeling and Western blotting experiments were performed using nuclear and cytoplasmic extracts of three human Philadelphia-negative leukemia/lymphoma cell lines (KG-1, HL-60, and Jurkat). Both methodologies showed that p160Bcr and p130Bcr localized to the cytoplasm, but the p130 form predominated in the nucleus. These results suggest that Bcr serves both nuclear and cytoplasmic functions, and that different forms of Bcr may be preferentially involved in these distinct activities.

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References

  1. Nowell PC, Hungerford DA . A minute chromosome in human chronic granulocytic leukemia Science 1960 132: 1497

    Google Scholar 

  2. Rowley JD . A new consistent chromosomal abnormality in chronic myelogenous leukemia identified by quinacrine fluorescence and Giemsa staining Nature 1973 243: 290–293

    Article  CAS  Google Scholar 

  3. Heisterkamp N, Groffen J, Stephenson JR, Spurr NK, Goodfellow PN, Solomon E, Carritt B, Bodmer WF . Chromosomal localization of human cellular homologues of two viral oncogenes Nature 1982 299: 747–749

    Article  CAS  Google Scholar 

  4. Kurzrock R, Gutterman JU, Talpaz M . The molecular genetics of Philadelphia chromosome-positive leukemias New Engl J Med 1988 319: 990–998

    Article  CAS  Google Scholar 

  5. Shtivelman E, Lifshitz B, Gale RP, Canaani E . Fused transcript of abl and bcr genes in chronic myelogenous leukaemia Nature 1985 315: 550–554

    Article  CAS  Google Scholar 

  6. Groffen J, Stephenson JR, Heisterkamp N, de Klein A, Bartram CR, Grosveld G . Philadelphia chromosomal breakpoints are clustered within a limited region, bcr, on chromosome 22 Cell 1984 36: 93–99

    Article  CAS  Google Scholar 

  7. Melo JV, Gordon DE, Cross NC, Goldman JM . The ABL-BCR fusion gene is expressed in chronic myeloid leukemia Blood 1993 81: 158–165

    CAS  PubMed  Google Scholar 

  8. Konopka JB, Watanabe SM, Witte ON . An alteration of the human c-Abl protein in K562 leukemia cells unmasks associated tyrosine kinase activity Cell 1984 37: 1035–1042

    Article  CAS  Google Scholar 

  9. Kloetzer W, Kurzrock R, Smith L, Talpaz M, Spiller M, Gutterman J, Arlinghaus R . The human cellular abl gene product in the chronic myelogenous leukemia cell line K562 has an associated tyrosine protein kinase activity Virology 1985 140: 230–238

    Article  CAS  Google Scholar 

  10. Kurzrock R, Shtalrid M, Romero P, Kloetzer W, Talpaz M, Trujillo JM, Blick M, Beran M, Gutterman JU . A novel c-Abl protein product in Philadelphia chromosome-positive leukemias Nature 1987 325: 631–635

    Article  CAS  Google Scholar 

  11. Arlinghaus RB . Multiple BCR-related gene products and their proposed involvement in ligand-induced signal transduction pathways Mol Carcinog 1992 5: 171–173

    Article  CAS  Google Scholar 

  12. Stam K, Heisterkamp N, Reynolds FH Jr, Groffen J . Evidence that the Phl gene encodes a 160 000-dalton phosphoprotein with associated kinase activity Mol Cell Biol 1987 7: 1955–1960

    Article  CAS  Google Scholar 

  13. Maru Y, Witte ON . The BCR gene encodes a novel serine/threonine kinase activity within a single exon Cell 1991 67: 459–468

    Article  CAS  Google Scholar 

  14. Ron D, Zanninni M, Lewis M, Wickner RB, Hunt LT, Graziani G, Tronick SR, Aaronson SA, Eva A . A region of proto-dbl essential for its transforming activity shows sequence similarity to a yeast cell cycle gene, CDC24, and the human breakpoint cluster gene, bcr New Biol 1991 3: 372–379

    CAS  PubMed  Google Scholar 

  15. Hart MJ, Eva A, Evans T, Aaronson SA, Cerione RA . Catalysis of guanine nucleotide exchange on the CDC42Hs protein by the DBL oncogene product Nature 1991 354: 311–314

    Article  CAS  Google Scholar 

  16. Adams JM, Houston H, Allen J, Lints T, Harvey R . The hematopoietically expressed vav proto-oncogene shares homology with the dbl GDP-GTP exchange factor, the bcr gene and a yeast gene (CDC24) involved in cytoskeletal organization Oncogene 1992 7: 611–618

    CAS  Google Scholar 

  17. Dhut S, Dorey EL, Horton MA, Ganesan TS, Young BD . Identification of two normal bcr gene products in the cytoplasm Oncogene 1988 3: 561–566

    CAS  PubMed  Google Scholar 

  18. Wetzler M, Talpaz M, Van Etten RA, Hirsh-Ginsberg C, Beran M, Kurzrock R . Subcellular localization of Bcr, Abl, and Bcr-Abl proteins in normal and leukemic cells and correlation of expression with myeloid differentiation J Clin Invest 1993 92: 1925–1939

    Article  CAS  Google Scholar 

  19. Wetzler M, Talpaz M, Yee G, Stass SA, Van Etten RA, Andreeff M, Goodacre AM, Kleine HD, Mahadevia RK, Kurzrock R . Cell cycle-related shifts in subcellular localization of BCR: association with mitotic chromosomes and with heterochromatin PNAS 1995 92: 3488–3492

    Article  CAS  Google Scholar 

  20. Dhut S, Chaplin T, Young BD . BCR-ABL and BCR proteins: biochemical characterization and localization Leukemia 1990 4: 745–750

    CAS  PubMed  Google Scholar 

  21. Amson RB, Marcelle C, Telerman A . Identification of a 130 kDa bcr-related gene product Oncogene 1989 4: 243–247

    CAS  PubMed  Google Scholar 

  22. Li WJ, Dreazen O, Kloetzer W, Gale RP, Arlinghaus RB . Characterization of BCR gene products in hematopoietic cells Oncogene 1989 4: 127–138

    CAS  PubMed  Google Scholar 

  23. Dignam JD, Lebovitz RM, Roeder RG . Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei Nucleic Acids Res 1983 11: 1475–1489

    Article  CAS  Google Scholar 

  24. Abmayr SM, Workman JL . DNA–protein interactions. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds) Current Protocols in Molecular Biology vol. 2: 5th edn Greene Publishing Associates and Wiley-Interscience: New York 1991 pp 12.0.1–12.9.4

    Google Scholar 

  25. Schaller MD, Borgman CA, Cobb BS, Vines RR, Reynolds AB, Parsons JT . pp125FAK a structurally distinctive protein-tyrosine kinase associated with focal adhesions PNAS 1992 89: 5192–5196

    Article  CAS  Google Scholar 

  26. Voncken JW, Hermien van Schaick H, Kaartinen V, Deemer K, Coates T, Landing B, Pattengale P, Dorseuil O, Bokoch GM, Groffen J, Heisterkamp N . Increased neutrophil respiratory burst in Bcr-null mutants Cell 1995 80: 719–728

    Article  CAS  Google Scholar 

  27. Reuther GW, Fu H, Cripe LD, Collier RJ, Pendergast AM . Association of the protein kinases c-Bcr and Bcr-Abl with proteins of the 14–3–3 family Science 1994 266: 129–133

    Article  CAS  Google Scholar 

  28. Braselmann S, McCormick F . BCR and RAF form a complex in vivo via 14–3–3 proteins EMBO J 1995 14: 4839–4848

    Article  CAS  Google Scholar 

  29. Bihn EA, Paul A-L, Wang SW, Erdos GW, Ferl RJ . Localization of 14–3–3 proteins in the nuclei of arabidopsis and maize Plant J 1997 12: 1439–1445

    Article  CAS  Google Scholar 

  30. Moore BW, Perez VJ . Specific acidic proteins of the nervous system. In: Carlson FD (ed.) Physiological and Biochemical Aspects of Nervous Intergration Prentice-Hall: Woods Hole, MA 1967 pp 343–359

    Google Scholar 

  31. Aitkin A . 14–3–3 proteins on the MAP Trends Biochem Sci 1995 20: 95–97

    Article  Google Scholar 

  32. Ford JC, Al-Khodairy F, Fotou E, Sheldrick KS, Griffiths DJF, Carr AM . 14–3–3 proteins homologs required for the DNA damage checkpoint in fission yeast Science 1994 265: 533–535

    Article  CAS  Google Scholar 

  33. Lopez-Girona A, Furnari B, Mondesert O, Russell P . Nuclear localization of Cdc25 is regulated by DNA damage and a 14–3–3 protein Nature 1999 397: 172–175

    Article  CAS  Google Scholar 

  34. Freed E, Symons M, MacDonald SG, McCormick F, Ruggieri R . Binding of 14–3–3 proteins to the protein kinase Raf and effects on its activation Science 1994 265: 1713–1716

    Article  CAS  Google Scholar 

  35. Irie K, Gotoh Y, Yashar BM, Errede B, Nishida E, Matsumoto K . Stimulatory effects of yeast and mammalian 14–3–3 proteins on the Raf protein kinase Science 1994 265: 1716–1719

    Article  CAS  Google Scholar 

  36. Fu H, Xia K, Pallas DC, Cui C, Conroy K, Narsimhan RP, Mamon H, Collier RJ, Roberts TM . Interaction of the protein kinase Raf-1 with 14–3–3 proteins Science 1994 266: 126–129

    Article  CAS  Google Scholar 

  37. Pendergast AM, Muller AJ, Havlik MH, Maru Y, Witte ON . BCR sequences essential for transformation by the BCR-ABL oncogene binds to the ABL SH2 regulatory domain in a nonphosphotyrosine-dependent manner Cell 1991 66: 161–171

    Article  CAS  Google Scholar 

  38. Muller AJ, Pendergast AM, Havlik MH, Puil L, Pawson T, Witte ON . A limited set of SH2 domains bind Bcr through a high-affinity phosphotyrosine-independent interaction Mol Cell Biol 1992 12: 5087–5093

    Article  CAS  Google Scholar 

  39. Liu J, Wu Y, Arlinghaus RB . Sequences within first exon of BCR inhibit the activated tyrosine kinase of c-Abl and the Bcr-Abl oncoprotein Cancer Res 1996 56: 5120–5124

    CAS  PubMed  Google Scholar 

  40. Liu J, Wu Y, Lu L, Arlinghaus RB . Inhibition of Bcr serine kinase by tyrosine phosphorylation Mol Cell Biol 1996 16: 998–1003

    Article  CAS  Google Scholar 

  41. Van Etten RA, 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 1989 58: 669–678

    Article  CAS  Google Scholar 

  42. Kharbanda S, Pandey P, Morris PL, Whang Y, Xu Y, Sawant S, Zhu L-J, Kumar N, Yuan Z-M, Weichselbaum R, Sawyers C, Pandita TK, Kufe D . Functional role for the c-Abl tyrosine kinase in meiosis I Oncogene 1998 16: 1773–1777

    Article  CAS  Google Scholar 

  43. Kipreos ET, Wang JYJ . Differential phosphorylation of c-Abl in cell cycle determined by cdc2 kinase and phosphatase activity Science 1990 248: 217–220

    Article  CAS  Google Scholar 

  44. Maru Y, Kobayashi T, Tanaka K, Shibuya M . Bcr binds to the xeroderma pigmentosum group B protein Biochem Biophys Res Comm 1999 260: 309–312

    Article  CAS  Google Scholar 

  45. Takeda N, Shibuya M, Maru Y . The Bcr-Abl oncoprotein potentially interacts with the xeroderma pigmentosum group B protein Proc Natl Acad Sci USA 1999 96: 203–207

    Article  CAS  Google Scholar 

  46. Frit P, Bergmann E, Egly J-M . Transcription factor IIH: a key player in the cellular response to DNA damage Biochimie 1999 81: 27–38

    Article  CAS  Google Scholar 

  47. Shi CS, Tuscano JM, Witte ON, Kehrl JH . GCKR links the Bcr-abl oncogene and Ras to the stress activated protein kinase pathway Blood 1999 93: 1338–1345

    CAS  PubMed  Google Scholar 

  48. McWhirter JR, Wang JY . Effect of Bcr sequences on the cellular function of the Bcr-Abl oncoprotein Oncogene 1997 15: 1625–1634

    Article  CAS  Google Scholar 

  49. Wu Y, Ma G, Lu D, Lin F, Xu HJ, Liu J, Arlinghaus RB . Bcr a negative regulator of the Bcr-Abl oncoprotein Oncogene 1999 18: 4416–4424

    Article  CAS  Google Scholar 

  50. Voncken JW, Kaartinen B, Groffen J, Hesterkamp N . Bcr/Abl associated leukemogenesis in bcr null mutant mice Oncogene 1998 16: 2029–2032

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported in part by an ACS Junior Faculty Research Award JFRA-655 (Meir Wetzler).

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Authors and Affiliations

  1. Department of Bioimmunotherapy, University of Texas MD Anderson Cancer Center, Houston, TX, USA

    E Laurent, M Talpaz & R Kurzrock

  2. Department of Medicine, Roswell Park Cancer Institute, Buffalo, NY, USA

    M Wetzler

  3. Department of Leukemia, University of Texas MD Anderson Cancer Center, Houston, TX, USA

    R Kurzrock

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  1. E Laurent
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  2. M Talpaz
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  3. M Wetzler
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  4. R Kurzrock
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Laurent, E., Talpaz, M., Wetzler, M. et al. Cytoplasmic and nuclear localization of the 130 and 160 kDa Bcr proteins. Leukemia 14, 1892–1897 (2000). https://doi.org/10.1038/sj.leu.2401923

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