Abstract
We have cloned cDNA for protein tyrosine phosphatase beta2, which had been implicated in erythroid differentiation of mouse erythroleukemia cells. Expression of cDNA constructs, in which beta2 cDNA is placed under the control of mouse metallothionein-I promoter, by ZnCl2 converted a significant portion (20 to 38%) of the cells to erythroid-like cells, which is 25-50% of the erythroid differentiation efficiency observed by conventional erythroid-inducing agents. Furthermore, introduction and expression of altered protein tyrosine phosphatase beta2 cDNA constructs designed to produce the enzyme lacking the phosphatase activity inhibited erythroid differentiation by 100-20%, depending upon the concentration of erythroid-inducing agents employed. These results strongly suggest that protein tyrosine phosphatase beta2 is involved in triggering erythroid differentiation in mouse erythroleukemia cells.
Highlights
Erythroleukemia cells have been the subject of extensive studies as one of the best models for terminal differentiation [1,2,3,4,5,6]
Introduction and expression of altered protein tyrosine phosphatase 2 cDNA constructs designed to produce the enzyme lacking the phosphatase activity inhibited erythroid differentiation by 100 –20%, depending upon the concentration of erythroid-inducing agents employed. These results strongly suggest that protein tyrosine phosphatase 2 is involved in triggering erythroid differentiation in mouse erythroleukemia cells
Upon exposure to inducing agents, mouse erythroleukemia (MEL)1 cells are converted to cells that exhibit all the characteristics of erythroid cells, including the loss of growth proficiency
Summary
Erythroleukemia cells have been the subject of extensive studies as one of the best models for terminal differentiation [1,2,3,4,5,6]. Upon exposure to inducing agents, mouse erythroleukemia (MEL) cells are converted to cells that exhibit all the characteristics of erythroid cells, including the loss of growth proficiency. Two intracellular protein factors implicated in MEL cell differentiation had characteristics of a tyrosine-phosphorylated protein and a PTPase, respectively [12]. We found that transcripts of two new PTPases, termed RIP and PTP2, were induced at a very early stage of differentiation [13]. We report cloning of a full-length cDNA for PTP2 and show that introduction and subsequent expression of the full-length PTP2 cDNA in MEL cells, but not RIP cDNA, convert a significant portion (20 –38%) of the cells to erythroid-like cells. Dominantnegative experiments using altered PTP2 cDNA constructs further suggested that PTP2 is involved in triggering MEL cell erythroid differentiation
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