title: Clonal variability in beta-globin mRNA content in an interleukin-3-dependent bone marrow cell line transfected with the erythropoietin receptor before and after stimulation with erythropoietin.
abstract: Unexpected clonal variability was observed in the content of beta-globin mRNA in erythropoietin receptor (EpoR)-transfected Ba/F3 cells before and after exposure to erythropoietin (Epo). Of 11 clones selected by virtue of G418 resistance and positive EpoR expression, 5 clones showed high levels of beta(major)-globin mRNA before Epo exposure, with subsequent Epo treatment causing little or no increase in globin mRNA. Five clones had undetectable levels of globin mRNA before Epo stimulation, and they did not accumulate globin mRNA when exposed to Epo, exhibiting resistance to the differentiation inducing action of Epo. Only one clone exhibited the expected phenotype, a low level of globin mRNA before exposure to Epo, and a significant Epo-dependent accumulation of globin mRNA. Phosphorylation of tyrosyl residues of the EpoR, Stat5, and JAK2 occurred upon Epo stimulation in clones representing each category. Furthermore, electrophoretic mobility shift assays using a Stat5 consensus sequence showed a difference in the nuclear binding component among these clones. These findings indicate that (1) the attainment of EpoR+ Ba/F3 clones with the anticipated sensitivity to both the growth and differentiation inducing actions of Epo is a rare event and (2) STAT5 transcription factors were differently activated by Epo in clones that differed in sensitivity to Epo.
authors.affiliation: Department of Pharmacology, Cancer Center, Yale University School of Medicine, New Haven, CT 06520, USA.
authors.forename: K
authors.initials: K
authors.lastname: Ishiguro
authors.forename: A C
authors.initials: AC
authors.lastname: Sartorelli
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001854:Bone Marrow Cells; D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002999:Clone Cells; D004268:DNA-Binding Proteins / Q000378:metabolism; D004920:Erythropoiesis / Q000187:drug effects; D004921:Erythropoietin / Q000494:pharmacology*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D005839:Gentamicins / Q000494:pharmacology*; D005914:Globins / Q000235:genetics*; D007377:Interleukin-3 / Q000494:pharmacology*; D051379:Mice; D008894:Milk Proteins; D012333:RNA, Messenger / Q000235:genetics; D017467:Receptors, Erythropoietin / Q000502:physiology*; D050799:STAT5 Transcription Factor; D015534:Trans-Activators / Q000378:metabolism; D014162:Transfection
chemical_list: D000900:Anti-Bacterial Agents; D002087:Butyrates; D004268:DNA-Binding Proteins; D005839:Gentamicins; D007377:Interleukin-3; D008894:Milk Proteins; D012333:RNA, Messenger; D017467:Receptors, Erythropoietin; D050799:STAT5 Transcription Factor; D015534:Trans-Activators; D020148:Butyric Acid; D004921:Erythropoietin; D005914:Globins; C010680:antibiotic G 418