Disrupted Signaling in a Mutant J2E Cell Line That Shows Enhanced Viability, but Does Not Proliferate or Differentiate, with Erythropoietin
Journal of Biological Chemistry(1996)
摘要
The immature erythroid J2E cell line proliferates and terminally differentiates following erythropoietin stimulation, In contrast, the mutant J2E-NR clone does not respond to erythropoietin by either proliferating or differentiating. Here we show that erythropoietin can act as a viability factor for both the J2E and J2E-NR lines, indicating that erythropoietin-initiated maturation is separable from the prevention of cell death. The inability of J2E-NR cells to mature in response to erythropoietin was not due to a defect in the erythropoietin receptor sequence, although surface receptor numbers were reduced, Both the receptor and Janus kinase 2 were phosphorylated after erythropoietin stimulation of J2E-NR cells, However, protein interactions with the erythropoietin receptor and Grb2 were restricted in the mutant cells. Subsequent investigation of several other signaling molecules exposed numerous alterations in J2E-NR cells; phosphorylation changes to phosphatidylinositol 3-kinase, phospholipase C gamma, p120 GAP, and mitogen-activated protein kinases (p42 and p44) observed in erythropoietin-stimulated J2E cells were not seen in the J2E-NR line. These data indicate that some pathways activated during erythropoietin-induced differentiation may not be essential for the prevention of apoptosis.
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关键词
cell line
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