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Ship in AML and Normal

The inositol 5-phosphatase SHIP is expressed as 145 and 135 kDa proteins in blood and bone marrow cells in vivo, whereas carboxyl-truncated forms of SHIP are generated by proteolytic cleavage in vitro

Leukemia volume 15, pages 112–120 (2001)Cite this article

Abstract

The inositol polyphosphate 5-phosphatase SHIP plays an important role in negative signalling in B cells and mast cells and in the down-regulation of cytokine receptor-mediated signals in myeloid cells. SHIP is expressed as a 145 kDa full-length protein and an isoform of 135 kDa due to alternative splicing. Additional smaller forms of SHIP which are truncated at the carboxy terminus have been described in bone marrow and peripheral blood mononuclear cells (PBMC). Our data demonstrate that human bone marrow cells and PBMC from healthy donors and patients with acute myeloid leukemia express the 145 kDa form of SHIP and low amounts of a 135 kDa form of SHIP in vivo whereas C-terminal-truncated SHIP proteins are generated by a PMSF-sensitive protease during the preparation of cell lysates in vitro. We have further characterized this protease and identified a proteolytic cleavage site in the human SHIP protein C-terminal to tryptophan residue 941. These data support a physiological role for the 145 and 135 kDa forms of SHIP in bone marrow and peripheral blood cells from normal donors and patients with acute myeloid leukemia.

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References

  1. Liu L, Damen JE, Cutler RL, Krystal G . Multiple Cytokines stimulate the binding of a common 145-kilodalton protein to Shc at the Grb2 recognition site of Shc Mol Cell Biol 1994 14: 6926–6935

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  2. Damen JE, Liu L, Rosten P, Humphries RK, Jefferson AB, Majerus PW, Krystal G . The 145-kDa protein induced to associate with Shc by multiple cytokines is an inositol tetraphosphate and phosphatidylinositol 3,4,5-trisphosphate 5-phosphatase Proc Natl Acad Sci USA 1996 93: 1689–1693

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  3. Kavanaugh WM, Pot DA, Chin SM, Deuter-Reinhard M, Jefferson AB, Norris FA, Masiarz FR, Cousens LS, Majerus PW, Williams LT . Multiple forms of an inositol polyphosphate 5-phosphatase form signaling complexes with Shc and Grb2 Curr Biol 1996 6: 438–445

    Article  CAS  PubMed  Google Scholar 

  4. Lioubin MN, Algate PA, Schickwann Tsai, Carlberg K, Aebersold R, Rohrschneider LR . p150Ship, a signal transduction molecule with inositol polyphosphate-5-phosphatase activity Genes Dev 1996 10: 1084–1095

    Article  CAS  PubMed  Google Scholar 

  5. Helgason CD, Damen JE, Rosten P, Grewal R, Sorenson P, Chappel SM, Borowski A, Jirik F, Krystal G, Humphries RK . Targeted disruption of SHIP leads to hematopoietic perturbations, lung pathology, and a shortened life span Genes Dev 1998 12: 1610–1620

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  6. Liu Q, Sasaki T, Kozieradzki I, Wakeham A, Itie A, Dumont DJ, Penninger JM . SHIP is a negative regulator of growth factor receptor-mediated PKB/Akt activation and myeloid cell survival Genes Dev 1999 13: 786–791

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  7. Liu Q, Shalaby F, Jones J, Bouchard D, Dumaont DJ . The SH2-Containing inositol polyphosphate 5-Phosphatase, SHIP, is expressed during hematopoesis and spermatogenesis Blood 1998 91: 2753–2759

    CAS  PubMed  Google Scholar 

  8. Geier SJ, Algate PA, Carlberg K, Flowers D, Friedmann C, Trask B, Rohrschneider LR . The human SHIP gene is differentially expressed in cell lineages of the bone marrow and blood Blood 1997 89: 1876–1885

    CAS  PubMed  Google Scholar 

  9. Damen JE, Liu L, Ware MD, Ermolaeva M, Majerus PW, Krystal G . Multiple forms of the SH2-containing inositol phosphatase, SHIP, are generated by c-terminal truncation Blood 1998 92: 1199–1205

    CAS  PubMed  Google Scholar 

  10. Lamkin TD, Walk SF, Liu L, Damen JE, Krystal G, Ravichandran KS . Shc interaction with Src homology 2 domain containing inositol phosphatase (SHIP) in vivo requires the Shc-phosphotyrosine binding domain and two specific phosphotyrosines on SHIP J Biol Chem 1997 272: 10396–10401

    Article  CAS  PubMed  Google Scholar 

  11. Kitamura T, Tange T, Terasawa T, Chiba S, Kuwaki T, Miyagawa K, Piao Y, Miyazono K, Urabe A, Takaku F . Establishment and characterization of a unique human cell line that proliferates dependently on GM-CSF, IL-3, or erythropoietin J Cell Physiol 1989 140: 323–334

    Article  CAS  PubMed  Google Scholar 

  12. Boettinger D, Anderson D, Dexter TM . Effect of src infection on long-term marrow cultures: increased self-renewal of hemopoietic progenitor cells without leukemia Cell 1984 36: 763–773

    Article  Google Scholar 

  13. Spooncer E, Heyworth CM, Dunn A, Dexter TM . Self renewal and differentiation of interleukin-3-independent multipotent stem cells are modulated by stromal cells and serum factors Differentiation 1986 31: 111–118

    Article  CAS  PubMed  Google Scholar 

  14. Dexter TM, Garland J, Scott D, Scolnick E, Metcalf D . Growth of factor-dependent hemopoietic precursor cell lines J Exp Med 1980 152: 1036–1047

    Article  CAS  PubMed  Google Scholar 

  15. Greenberger JS, Eckner R, Ostertag W, Colleta B, Boshetti S, Nagasawa H, Karpas A, Weichselbaum R, Moloney W . Release of spleen focus-forming virus (SFFV) from differentiation inducible promyelocytic leukemia cell lines transformed in vitro by Friend leukemia virus Virology 1980 105: 425–435

    Article  CAS  PubMed  Google Scholar 

  16. Ihle JN . Biochemical and biological properties of interleukin-3: a lymphokine mediating the differentiation of a lineage of cells which includes prothymocytes and mast-like cells Contemp Topics Mol Immunol 1985 10: 93–119

    CAS  Google Scholar 

  17. Meyer J, Jücker M, Ostertag W, Stocking C . Carboxyl-truncated STAT5b is generated by a nucleus-associated serine protease in early hematopoietic progenitors Blood 1998 91: 1901–1908

    CAS  PubMed  Google Scholar 

  18. Fehse B, Uhde A, Fehse N, Eckert HG, Clausen J, Rüger R, Koch S, Ostertag W, Zander AR, Stockschläder M . Selective immunoaffinity-based enrichment of CD34+ cells transduced with retroviral vectors containing an intracytoplasmatically truncated version of the human low-affinity nerve growth factor receptor (deltaLNGFR) gene Hum Gene Ther 1997 8: 1815–1824

    Article  CAS  PubMed  Google Scholar 

  19. Jücker M, Schaadt M, Diehl V, Poppema S, Jones D, Tesch H . Heterogeneous expression of proto-oncogenes in Hodgkin's disease derived cell lines Hematol Oncol 1990 8: 191–204

    Article  PubMed  Google Scholar 

  20. Jücker M, Abts H, Li W, Schindler R, Merz H, Günther A, von Kalle C, Schaadt M, Diamantstein T, Feller AC, Krueger GRF, Diehl V, Blankenstein T, Tesch H . Expression of interleukin-6 and interleukin-6 receptor in Hodgkin's disease Blood 1991 77: 2413–2418

    PubMed  Google Scholar 

  21. Areces LB, Deello Sbarba P, Jücker M, Stanley ER, Feldman RA . Functional specificity of cytoplasmic and transmembrane tyrosine kinases: identification of 130 and 75 kda substrates of c-fps/fes tyrosine kinase in macrophages Mol Cell Biol 1994 14: 4606–4615

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  22. Jücker M, Feldman RA . Identification of a new adapter proteinthat may link the common β subunit of the receptor for granulocyte/macrophage colony-stimulating factor, interleukin (IL)-3, and IL-5 to phosphatidylinositol 3-kinase J Biol Chem 1995 270: 27817–27822

    Article  PubMed  Google Scholar 

  23. Ware MD, Rosten P, Damen JE, Liu L, Humphries RK, Krystal G . Cloning and characterization of human SHIP, the 145-kD inositol 5-phsophatase that associates with Shc after cytokine stimulation Blood 1996 88: 2833–2840

    CAS  PubMed  Google Scholar 

  24. Schulze C, Heukeshoven J . Average and maximum charge states of arginine-containing dendrimer-like peptide ions formed by electrospray ionization Eur Mass Spectrom 1998 4: 133–139

    Article  CAS  Google Scholar 

  25. Müller D, Schulze C, Baumeister H, Buck F, Richter D . Rat insulin-degrading enzyme: cleavage pattern of the natriuretic peptide hormones ANP, BNP, and CNP revealed by HPLC and mass spectrometry Biochem 1992 31: 11138–11143

    Article  Google Scholar 

  26. Lucas DM, Rohrschneider LR . A novel spliced form of SH2-containing inositol phosphatase is expressed during myeloid development Blood 1999 93: 1922–1933

    CAS  PubMed  Google Scholar 

  27. Yi T, Mui AL, Krystal G, Ihle JN . Hematopoietic cell phosphatase associates with the interleukin-3 (IL-3) receptor beta chain and down-regulates IL-3-induced tyrosine phosphorylation and mitogenesis Mol Cell Biol 1993 13: 7577–7586

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  28. Yoshimura A, Ohkubo T, Kiguchi T, Jenkins NA, Gilbert DJ, Copeland NG, Hara T, Miyajima A . A novel cytokine-inducible gene CIS encodes an SH2-containing protein that binds to tyrosine-phosphorylated interleukin 3 and erythropoietin receptors EMBO J 1995 14: 2816–2826

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  29. Matsumoto A, Masuhara M, Mitsui K, Yokouchi M, Ohtsubo M, Misawa H, Miyajima A, Yoshimura A . CIS, a cytokine inducible SH2 protein, is a target of the JAK-STAT5 pathway and modulates STAT5 activation Blood 1997 89: 3148–3154

    CAS  PubMed  Google Scholar 

  30. Endo TA, Masuhara M, Yokouchi M, Suzuki R, Sakamoto H, Mitsui K, Matsumoto A, Tanimura S, Ohtsubo M, Misawa H, Miyazaki T, Leonor N, Taniguchi T, Fujita T, Kanakura Y, Komiya S, Yoshimura A . A new protein containing an SH2 domain that inhibits JAK kinases Nature 1997 387: 921–924

    Article  CAS  PubMed  Google Scholar 

  31. Starr R, Willson TA, Viney EM, Murray LJL, Rayner JR, Jenkins BJ, Gonda TJ, Alexander WS, Metcalf D, Nicola NA, Hilton DJ . A family of cytokine-inducible inhibitors of signalling Nature 1997 387: 917–921

    Article  CAS  PubMed  Google Scholar 

  32. Naka T, Narazaki M, Hirata M, Matsumoto T, Minamoto S, Aono A, Nishimoto N, Kajita T, Taga T, Yoshizaki K, Akira S, Kishimoto T . Structure and function of a new STAT-induced STAT inhibitor Nature 1997 387: 924–928

    Article  CAS  PubMed  Google Scholar 

  33. Ono M, Bolland S, Tempst P, Ravetsch JV . Role of the inositol phosphatase SHIP in negative regulation of the immune system by the receptor FcgRIIB Nature 1996 383: 263–266

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We would like to thank W Fiedler, G Schuch and DK Hossfeld from the Department of Hematology/Oncology, University Hospital Eppendorf, Hamburg, for providing leukemia cells and recombinant human GM-CSF and C Laker for preparing murine bone marrow cells. We are greatful to L Rohrschneider for providing the murine SHIP cDNA. This work was supported by grants from the Deutsche Forschungsgemeinschaft (JU255/2–1) and Thyssen Stiftung (CS).

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Authors and Affiliations

  1. Abteilung für Zelluläre Signaltransduktion, Institut für Medizinische Biochemie und Molekularbiologie, Universitäts-Krankenhaus Eppendorf, Universität Hamburg, Germany

    S Horn, S Poll & M Jücker

  2. Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie an der Universität Hamburg, Germany

    J Meyer, J Heukeshoven & C Stocking

  3. Knochenmark-Transplantationszentrum, Universitäts-Krankenhaus Eppendorf, Germany

    B Fehse

  4. Zentrum für Molekulare Neurobiologie, Universität Hamburg, Hamburg, Germany

    C Schulze

  5. Biochemie II der Universität Bielefeld, Bielefeld, Germany

    S Li & J Frey

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Horn, S., Meyer, J., Heukeshoven, J. et al. The inositol 5-phosphatase SHIP is expressed as 145 and 135 kDa proteins in blood and bone marrow cells in vivo, whereas carboxyl-truncated forms of SHIP are generated by proteolytic cleavage in vitro. Leukemia 15, 112–120 (2001). https://doi.org/10.1038/sj.leu.2401990

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