EXPRESSION of ACTIVATED of MAP KINASE KINASE IS SUFFICIENT for MEGAKARYOCYTIC DIFFERENTIATION of CMK CELLS

Allen S. Melemed, John Rvder, Terry Vik, David A. Williams

Research output: Contribution to journalArticle

Abstract

Activation of the MAP kinase pathway has been shown to be important in determining cell fate in non-hematopoietic cells. The activation of the MAP kinase pathway through the expression of constitutively activated MAP kinase kinase (MAPKK) induces differentiation in PC 12 cells, a pheochromocytoma cell line. Conversely, expression of constitutively activated MAPKK in NIH 3T3 cells causes transformation. The role of the MAP kinase pathway with regards to differentiation or proliferation has not been directly examined in hematopoietic cell lines. We examined the role of the MAP kinase pathway by the stable transfection of a human megakaryocytic cell line, CMK, with wild type and constitutively activated MAPKK. Constitutively activated MAPKK was subcloned into a eukaryotic expression plasmid, pcDNA3, and electroporated into CMK cells. The resultant stablely transfected constitutively activated MAPKK clones showed morphologic changes consistent with differentiation including increased size, decreased nuclear to cytoplasmic ratio, and multinucleation. These clones expressed mature megakaryocytic cell surface markers, CD 41a and CD 61 by FACScan analysis. The clones transfected with the cDNA encoding the activated MAPKK showed increased MAP kinase activity compared to clones expressing wild type MAPKK. We conclude that the expression of constitutively activated MAPKK is sufficient to induce megakaryocytic differentiation in the CMK hematopoietic cell line.

Original languageEnglish
Pages (from-to)1109
Number of pages1
JournalExperimental Hematology
Volume24
Issue number9
StatePublished - 1996

Fingerprint

Mitogen-Activated Protein Kinase Kinases
Phosphotransferases
Clone Cells
Cell Line
NIH 3T3 Cells
PC12 Cells
Transfection
Plasmids
Complementary DNA

ASJC Scopus subject areas

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

Cite this

EXPRESSION of ACTIVATED of MAP KINASE KINASE IS SUFFICIENT for MEGAKARYOCYTIC DIFFERENTIATION of CMK CELLS. / Melemed, Allen S.; Rvder, John; Vik, Terry; Williams, David A.

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

Research output: Contribution to journalArticle

Melemed, Allen S. ; Rvder, John ; Vik, Terry ; Williams, David A. / EXPRESSION of ACTIVATED of MAP KINASE KINASE IS SUFFICIENT for MEGAKARYOCYTIC DIFFERENTIATION of CMK CELLS. In: Experimental Hematology. 1996 ; Vol. 24, No. 9. pp. 1109.
@article{2ce98ddfb29740eb8cffc2922378cce6,
title = "EXPRESSION of ACTIVATED of MAP KINASE KINASE IS SUFFICIENT for MEGAKARYOCYTIC DIFFERENTIATION of CMK CELLS",
abstract = "Activation of the MAP kinase pathway has been shown to be important in determining cell fate in non-hematopoietic cells. The activation of the MAP kinase pathway through the expression of constitutively activated MAP kinase kinase (MAPKK) induces differentiation in PC 12 cells, a pheochromocytoma cell line. Conversely, expression of constitutively activated MAPKK in NIH 3T3 cells causes transformation. The role of the MAP kinase pathway with regards to differentiation or proliferation has not been directly examined in hematopoietic cell lines. We examined the role of the MAP kinase pathway by the stable transfection of a human megakaryocytic cell line, CMK, with wild type and constitutively activated MAPKK. Constitutively activated MAPKK was subcloned into a eukaryotic expression plasmid, pcDNA3, and electroporated into CMK cells. The resultant stablely transfected constitutively activated MAPKK clones showed morphologic changes consistent with differentiation including increased size, decreased nuclear to cytoplasmic ratio, and multinucleation. These clones expressed mature megakaryocytic cell surface markers, CD 41a and CD 61 by FACScan analysis. The clones transfected with the cDNA encoding the activated MAPKK showed increased MAP kinase activity compared to clones expressing wild type MAPKK. We conclude that the expression of constitutively activated MAPKK is sufficient to induce megakaryocytic differentiation in the CMK hematopoietic cell line.",
author = "Melemed, {Allen S.} and John Rvder and Terry Vik and Williams, {David A.}",
year = "1996",
language = "English",
volume = "24",
pages = "1109",
journal = "Experimental Hematology",
issn = "0301-472X",
publisher = "Elsevier Inc.",
number = "9",

}

TY - JOUR

T1 - EXPRESSION of ACTIVATED of MAP KINASE KINASE IS SUFFICIENT for MEGAKARYOCYTIC DIFFERENTIATION of CMK CELLS

AU - Melemed, Allen S.

AU - Rvder, John

AU - Vik, Terry

AU - Williams, David A.

PY - 1996

Y1 - 1996

N2 - Activation of the MAP kinase pathway has been shown to be important in determining cell fate in non-hematopoietic cells. The activation of the MAP kinase pathway through the expression of constitutively activated MAP kinase kinase (MAPKK) induces differentiation in PC 12 cells, a pheochromocytoma cell line. Conversely, expression of constitutively activated MAPKK in NIH 3T3 cells causes transformation. The role of the MAP kinase pathway with regards to differentiation or proliferation has not been directly examined in hematopoietic cell lines. We examined the role of the MAP kinase pathway by the stable transfection of a human megakaryocytic cell line, CMK, with wild type and constitutively activated MAPKK. Constitutively activated MAPKK was subcloned into a eukaryotic expression plasmid, pcDNA3, and electroporated into CMK cells. The resultant stablely transfected constitutively activated MAPKK clones showed morphologic changes consistent with differentiation including increased size, decreased nuclear to cytoplasmic ratio, and multinucleation. These clones expressed mature megakaryocytic cell surface markers, CD 41a and CD 61 by FACScan analysis. The clones transfected with the cDNA encoding the activated MAPKK showed increased MAP kinase activity compared to clones expressing wild type MAPKK. We conclude that the expression of constitutively activated MAPKK is sufficient to induce megakaryocytic differentiation in the CMK hematopoietic cell line.

AB - Activation of the MAP kinase pathway has been shown to be important in determining cell fate in non-hematopoietic cells. The activation of the MAP kinase pathway through the expression of constitutively activated MAP kinase kinase (MAPKK) induces differentiation in PC 12 cells, a pheochromocytoma cell line. Conversely, expression of constitutively activated MAPKK in NIH 3T3 cells causes transformation. The role of the MAP kinase pathway with regards to differentiation or proliferation has not been directly examined in hematopoietic cell lines. We examined the role of the MAP kinase pathway by the stable transfection of a human megakaryocytic cell line, CMK, with wild type and constitutively activated MAPKK. Constitutively activated MAPKK was subcloned into a eukaryotic expression plasmid, pcDNA3, and electroporated into CMK cells. The resultant stablely transfected constitutively activated MAPKK clones showed morphologic changes consistent with differentiation including increased size, decreased nuclear to cytoplasmic ratio, and multinucleation. These clones expressed mature megakaryocytic cell surface markers, CD 41a and CD 61 by FACScan analysis. The clones transfected with the cDNA encoding the activated MAPKK showed increased MAP kinase activity compared to clones expressing wild type MAPKK. We conclude that the expression of constitutively activated MAPKK is sufficient to induce megakaryocytic differentiation in the CMK hematopoietic cell line.

UR - http://www.scopus.com/inward/record.url?scp=33748613495&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33748613495&partnerID=8YFLogxK

M3 - Article

VL - 24

SP - 1109

JO - Experimental Hematology

JF - Experimental Hematology

SN - 0301-472X

IS - 9

ER -