Analysis of functional domains of the human tpo receptor encoded by C-MPL

M. Takatoku, N. Komateu, R. Shimizu, Takashi Kato, H. Miyazakis, Y. Miuräï

Research output: Contribution to journalArticle

Abstract

c-Mpl, a member of the hematopoietic cytokine receptor family, is the receptor for thrombopoietin (TPO). To establish a model system for analyzing functional domains of c-Mpl, we searched for a cell line not expressing endogenous c-Mpl. Previously we reported that c-mpl mRNA could not be detected in a human EPO-responsive cell line, UT7/EPO (Blood 82:456,1993; Blood in press). This was confirmed by the reverse transcriptase-polymerase chain reaction (RT-PCR) method. Based on these observations, we prepared an expression vector containingfulllengthc-mp/ cDNA and transfected it into the UT-7/EPO cells. We isolated 3 clones and examined whether these clones exogenously expressed c-Mpl on the surface of the cells by FACS analysis with an anti-c-Mpl antibody. All these transfectants expressed c-Mpl and grew in response to TPO. Moreover, after incubation with TPO for one week, some of the cells became large and multinucleated, and expressed platelet factor-4 mRNA, suggesting that the TPO signalling pathway mediated through c-Mpl is involved in not only the proliferation but also megakaryocytic differentiation of the transfectants. Therefore, this model system may be useful for analyzing the functional domains of human c-Mpl. We are now preparing several kinds of deletion mutants.

Original languageEnglish
Pages (from-to)1109
Number of pages1
JournalExperimental Hematology
Volume24
Issue number9
Publication statusPublished - 1996
Externally publishedYes

Fingerprint

Thrombopoietin
Clone Cells
Thrombopoietin Receptors
Platelet Factor 4
Cell Line
Messenger RNA
Cytokine Receptors
Reverse Transcriptase Polymerase Chain Reaction
Complementary DNA

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

Cite this

Takatoku, M., Komateu, N., Shimizu, R., Kato, T., Miyazakis, H., & Miuräï, Y. (1996). Analysis of functional domains of the human tpo receptor encoded by C-MPL. Experimental Hematology, 24(9), 1109.

Analysis of functional domains of the human tpo receptor encoded by C-MPL. / Takatoku, M.; Komateu, N.; Shimizu, R.; Kato, Takashi; Miyazakis, H.; Miuräï, Y.

In: Experimental Hematology, Vol. 24, No. 9, 1996, p. 1109.

Research output: Contribution to journalArticle

Takatoku, M, Komateu, N, Shimizu, R, Kato, T, Miyazakis, H & Miuräï, Y 1996, 'Analysis of functional domains of the human tpo receptor encoded by C-MPL', Experimental Hematology, vol. 24, no. 9, pp. 1109.
Takatoku M, Komateu N, Shimizu R, Kato T, Miyazakis H, Miuräï Y. Analysis of functional domains of the human tpo receptor encoded by C-MPL. Experimental Hematology. 1996;24(9):1109.
Takatoku, M. ; Komateu, N. ; Shimizu, R. ; Kato, Takashi ; Miyazakis, H. ; Miuräï, Y. / Analysis of functional domains of the human tpo receptor encoded by C-MPL. In: Experimental Hematology. 1996 ; Vol. 24, No. 9. pp. 1109.
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