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 language | English |
|---|---|
| Pages (from-to) | 1109 |
| Number of pages | 1 |
| Journal | Experimental Hematology |
| Volume | 24 |
| Issue number | 9 |
| Publication status | Published - 1996 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Cancer Research
- Cell Biology
- Genetics
- Hematology
- Oncology
- Transplantation
Cite this
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 journal › Article
}
TY - JOUR
T1 - Analysis of functional domains of the human tpo receptor encoded by C-MPL
AU - Takatoku, M.
AU - Komateu, N.
AU - Shimizu, R.
AU - Kato, Takashi
AU - Miyazakis, H.
AU - Miuräï, Y.
PY - 1996
Y1 - 1996
N2 - 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.
AB - 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.
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M3 - Article
VL - 24
SP - 1109
JO - Experimental Hematology
JF - Experimental Hematology
SN - 0301-472X
IS - 9
ER -