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Volume 271, Number 44, Issue of November 1, 1996 pp. 27432-27437
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

Interleukin-3 (IL-3) Inhibits Erythropoietin-induced Differentiation in Ba/F3 Cells via the IL-3 Receptor alpha  Subunit

(Received for publication, February 14, 1996, and in revised form, July 10, 1996)

Jana Krosl Dagger , Jacqueline E. Damen Dagger , Gerald Krystal Dagger § and R. Keith Humphries Dagger par

From the Dagger  Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, British Columbia V5Z 1L3, Canada and the Departments of § Pathology and par  Medicine, University of British Columbia, Vancouver, British Columbia V6T 1Z1, Canada

Introduction of erythropoietin receptors (EpoRs) into the interleukin-3 (IL-3)-dependent murine hemopoietic cell line, Ba/F3, enables these cells to not only proliferate, after an initial lag in G1, but also to increase beta -globin mRNA levels in response to erythropoietin (Epo). With IL-3 and Epo costimulation, IL-3-induced signaling appears to be dominant since no increase in beta -globin mRNA occurs. Differentiation and proliferation signals may be uncoupled since EpoRs lacking all eight intracellular tyrosines were compromised in proliferative signaling but retained erythroid differentiation ability. Intriguingly, a chimeric receptor of the extracellular domain of the EpoR and the transmembrane and intracellular domains of IL-3Rbeta IL-3 chain (EpoR/IL-3Rbeta IL-3) was capable of Epo-induced proliferative and differentiating signaling, suggesting either the existence of a second EpoR subunit responsible for differentiation or that the alpha  subunit of the IL-3 receptor (IL-3R) prevents it. Arguing against the former, a truncated EpoR lacking an intracellular domain was incapable of promoting proliferation or differentiation. An EpoR/IL-3Ralpha chimera, in contrast, was capable of transmitting a weak Epo-induced proliferative signal but failed to stimulate accumulation of beta -globin mRNA. Most significantly, coexpression of the EpoR/IL-3Ralpha chimera with either EpoR/IL-3Rbeta or wild-type EpoRs suppressed Epo-induced beta -globin mRNA accumulation. Taken together, these results suggest an active role for the IL-3Ralpha subunit in inhibiting EpoR-specific differentiating signals.


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Copyright © 1996 by the American Society for Biochemistry and Molecular Biology.