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1992 Fiscal Year Final Research Report Summary

Analysis of signal transduction mechanism through a novel tyrosine kinase receptor

Research Project

Project/Area Number 03454521
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Hematology
Research InstitutionThe University of Tokyo

Principal Investigator

HIRAI Hisamaru  Faculty of Medicine, University of Tokyo, Lecturer, 医学部(病), 講師 (90181130)

Co-Investigator(Kenkyū-buntansha) CHIBA Shigeru  Faculty of Medicine, University of Tokyo, Assistant Prof., 医学部(病), 助手
TOYOSHIMA Hideo  Faculty of Medicine, University of Tokyo, Assistant Prof., 医学部(病), 助手 (20197966)
MANO Hiroyuki  Faculty of Medicine, University of Tokyo, Assistant Prof., 医学部(病), 助手 (90240704)
Project Period (FY) 1991 – 1992
Keywordsltk protein / Phosphorylation / Receptor-type protein tyrosine kinase / Alternative splicing
Research Abstract

Previously, we cloned an ltk cDNA isolated from a human erythroleukemia cell line K562 cDNA library by low stringency hybridization with a c-fms probe. To obtain a full length cDNA, a cDNA library of human placenta was screened, and five positive clones were isolated. One clone (P2) contains the longest open reading frame which encodes a putative tyrosine kinase receptor of 864 amino acids. The analysis of the five clones revealed the existence of at least three spieces of cDNA clones. One cDNA predicts a protein of 221 amino acids and encodes a soluble type of receptor which can be secreted from the cell. Another cDNA clone (P3) has an alternative insert just downstream of the transmembrane domain and thus produce in-frame stop codon between the transmembrane domain and the tyrosine kinase domain, suggesting that this clone encodes a receptor protein lacking the kinase domain. We transfected expression plasmids containing various ltk cDNA clones to COS cells, and cell lysates were sub … More jected to western blot analysis. Proteins of approximately 100kD and 50kD were detected by the monoclonal antibody against the extracellular domain in the cell lysates transfected with P2 and P3, respectively. Both products are shown in double bands which may result from posttranslational modification. These results suggest that these cDNAs do not represent immature unspliced pre-mRNAs. Immune complexes from lysates of COS cells expressing the full ltk cDNA clone were subjected to in vitro kinase assay using [gamma-^<32>P]ATP. Phosphorylation of 100kD, 140kD, 85kD and 45kD proteins was detected in the experiment, suggesting that ltk protein has protein kinase activity and phosphorylates itself and other associated proteins. We have detected PLC-gamma1, GAP, PI3-kinase, and c-raf protein in the immune complex precipitated by anti-ltk antibody. In Northern blot analysis of 35 human malignancies, ltk is preferentially expressed in leukemias (10 out of 18 cases) with no cell lineage specificity, but none of 17 nonleukemic neoplasms expressed ltk gene. Less

  • Research Products

    (20 results)

All Other

All Publications (20 results)

  • [Publications] Sugimoto K et al.: "Freguent mutatious in the p53 gene in human myeloid leukemia cell lines." Blood. 79. 2378-2383 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hanazono.Y.et al: "Erythnopoietin induces throsine phosphorylation and kinase activity of the c-fps/fes proto-oncogene product in human erythropoietin-responsive cells." Blood.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toyoshima H et al.: "Differently spliced cDNAs of human ltk receptor tyrosine leinase predict receptor proteins with and without tyrosine leinase sowain and a sdubli receptor protein." Proc.Nael.Acad.Sci.USA.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sugimoto K et al.: "Mutations of the p53 gene en myelodysplastic syndromes and MDS-derived leukemia." Blood.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hanazono Y et al.: "c-fps/fes protein-tyrosine kinase is implicated in a signaling pathway criggered by granulocyte-macrophage colony-stimulating factor and IL-3." EMBO J.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sasaki K et al.: "Coordinate expression of the α and β chains of human. graw locyte-macrophege colony-stimulating factor receptor confers ligand-induced morphological transformation in mouse fibroblasts." J.Biol Chem.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sugimoto K etal: "Mutations of the p53 gene in lymphoid leukemia" Blood. 77. 1153-1156 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirai H et al: "Analysis of ph1-positive leukemia by PCR" Jpn.Clinical Hematol.32. 319-326 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Chiba S et al: "Establishment and erythroid differentiation of a cytokine-dependent human leukemic cell line F-36:Apasental line requining granulocyte-macrophage colony stimulating factor or interleukin-3 and a subline reqwining enjthropoietin" Blood. 78. 2261-2268 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kozutsumi et al: "Identification of the human ltk gene product in placeuta and hewatopoietic cell lines." Biochem.Biophys.Res.Commuu.

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Sugimoto K, Toyoshima H, Sakai R, Miyagawa K, Hagiwara K, Hirai H, Ishikawa F, Takaku F.: "Mutations of the p53 gene in lymphoid leukemia." Blood. 77. 1153-1156 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hirai H, Ishikawa F.: "Analysis of Ph1-positive leukemia by PCR." Jpn. Clinical Hematol. 32. 319-326 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Chiba S, Takaku F, Tange T, Shibuya K, Misawa C, Sasaki K, Miyagawa K, Yazaki Y, Hirai H.: "Establishment and erythroid differentiation of a cytokine-dependent human leukemic cell line F-36: A parental line requiring granulocyte-macrophage colony- stimulating factor or interleukin-3, and a subline requiring erythropoietin." Blood. 78. 2261-2268 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sugimoto K, Toyoshima H, Sakai R, Miyagawa K, Hagiwara K, Ishikawa F, Takaku F, Yazaki Y, Hirai H.: "Frequent mutations in the p53 gene in human myeloid leukemia cell lines." Blood. 79. 2378-2383 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kozutsumi H, Toyoshima H, Hagiwara K, Furuya A, Mioh H, Hanai N, Yazaki Y, Hirai H.: "Identification of the human ltk gene product in placenta and hematopoietic cell lines." Biochem. Biophys. Res. Commun.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toyoshima H, Kozutsumi H, Maru Y, Hagiwara K, Furuya A, Mioh H, Hanai N, Takaku F, Yazaki Y, Hirai H.: "Differently spliced cDNAs of human ltk receptor tyrosine kinase predict receptor proteins with and without tyrosine kinase domain and a soluble receptor protein." Proc. Natl. Acad. Sci. USA.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sugimoto K, Hirono N, Toyoshima H, Chiba S, Mano H, Takaku F, Yazaki Y, Hirai H.: "Mutations of the p53 gene in myelodysplastic syndromes and MDS-derived leukemia." Blood.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hanazono Y, Chiba S, Sasaki K, Mano H, Miyajima A, Arai K, Yazaki Y, Hirai H.: "c-fpslfes protein-tyrosine kinase is implicated in a signaling pathway triggered by granulocyte-macrophage colony-stimulating factor and interleukin-3." EMBO J.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hanazono Y, Chiba S, Sasaki K, Mano H, Yazaki Y, Hirai H.: "Erythropoietin induces tyrosine phosphorylation and kinase activity of the c-fpslfes proto-oncogene product in human erythropoietin-responsive cells." Blood.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sasaki K, Chiba S, Hanazono Y, Mano H, Yazaki Y, Hirai H.: "Coordinate expression of the a and b chains of human granolocyte-macrophage colony-stimulating factor receptor confers ligand-induced morphological transformation in mouse fibroblasts." J. Biol. Chem.

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1994-03-24  

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