title: STAT5 activation correlates with erythropoietin receptor-mediated erythroid differentiation of an erythroleukemia cell line.
abstract: Interaction between erythropoietin (EPO) and its membrane receptor induces the proliferation and differentiation of erythroid progenitors. EPO has been shown to activate the JAK2-STAT5 pathway in various hematopoietic cell lines, although the physiological role of this pathway is unclear. We have previously shown that epidermal growth factor activates a chimeric receptor bearing the extracellular domain of the epidermal growth factor receptor linked to the cytoplasmic domain of the EPO receptor, resulting in proliferation of interleukin-3-dependent hematopoietic cells and erythroid differentiation (globin synthesis) of EPO-responsive erythroleukemia cells. In the present study, we introduced various deletion and tyrosine to phenylalanine substitution in the cytoplasmic domain of the chimeric receptor and expressed these mutant chimeras in an EPO-responsive erythroleukemia cell line, ELM-I-1. Mutant chimeric receptors retaining either Tyr343 or Tyr401 could activate STAT5, judged by tyrosine-phosphorylation of STAT5 and induction of CIS, a target gene of STAT5. These mutants were able to induce erythroid differentiation. However, a chimeric receptor containing both Y343F and Y401F mutations could not activate STAT5 nor induce erythroid differentiation. Thus, Tyr343 or Tyr401 of the EPO receptor are independently necessary for erythroid differentiation as well as STAT5 activation. Moreover, exogenous expression of dominant-negative STAT5 suppressed EPO-dependent erythroid differentiation. These findings suggest that STAT5 plays an important role in erythroid differentiation through the EPO receptor cytoplasmic domain.
authors.affiliation: Institute of Life Science, Aikawamachi 2432-3 Kurume 839, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Iwatsuki
authors.forename: T
authors.initials: T
authors.lastname: Endo
authors.forename: H
authors.initials: H
authors.lastname: Misawa
authors.forename: M
authors.initials: M
authors.lastname: Yokouchi
authors.forename: A
authors.initials: A
authors.lastname: Matsumoto
authors.forename: M
authors.initials: M
authors.lastname: Ohtsubo
authors.forename: K J
authors.initials: KJ
authors.lastname: Mori
authors.forename: A
authors.initials: A
authors.lastname: Yoshimura
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D018076:DNA, Complementary / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000378:metabolism*; D015672:Erythroid Precursor Cells / Q000166:cytology*; D005622:Friend murine leukemia virus; D053614:Janus Kinase 2; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism; D008894:Milk Proteins; D016297:Mutagenesis, Site-Directed; D010649:Phenylalanine / Q000378:metabolism; D011487:Protein Conformation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins; D017467:Receptors, Erythropoietin / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D050799:STAT5 Transcription Factor; D015534:Trans-Activators / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D008894:Milk Proteins; D011518:Proto-Oncogene Proteins; D017467:Receptors, Erythropoietin; D011993:Recombinant Fusion Proteins; D050799:STAT5 Transcription Factor; D015534:Trans-Activators; D014443:Tyrosine; D010649:Phenylalanine; D011505:Protein-Tyrosine Kinases; D053614:Janus Kinase 2