title: Transcription factor erythroid Krüppel-like factor (EKLF) is essential for the erythropoietin-induced hemoglobin production but not for proliferation, viability, or morphological maturation.
abstract: The erythroid Krüppel-like factor (EKLF) is essential for the transcription of betamaj globin in erythroid cells. We show here that RNA for this transcription factor did not alter during erythropoietin-induced differentiation of J2E cells; however, EKLF protein content decreased and was inversely related to globin production. This unexpected result was also observed during chemically induced maturation of two murine erythroleukemia cell lines. To explore the role of EKLF in erythroid terminal differentiation, an antisense EKLF construct was introduced into J2E cells. As a consequence EKLF RNA and protein levels fell by approximately 80%, and the cells were unable to manufacture hemoglobin in response to erythropoietin. The failure to produce hemoglobin was due to reduced transcription of not only globin genes but also key heme enzyme genes. However, numerous other genes, including several erythroid transcription factors, were unaffected by the decrease in EKLF. Although hemoglobin synthesis was severely impaired with depleted EKLF levels, morphological maturation in response to erythropoietin continued normally. Moreover, erythropoietin-induced proliferation and viability were unaffected by the decrease in EKLF levels. We conclude that EKLF affects a specific set of genes, which regulates hemoglobin production and has no obvious effect on morphological changes, cell division, or viability in response to erythropoietin.
authors.affiliation: Laboratory for Cancer Medicine, Department of Biochemistry, Level 6, MRF Building, Rear, 50 Murray Street, Perth, Western Australia 6001 Australia.
authors.forename: A
authors.initials: A
authors.lastname: Spadaccini
authors.forename: P A
authors.initials: PA
authors.lastname: Tilbrook
authors.forename: M K
authors.initials: MK
authors.lastname: Sarna
authors.forename: M
authors.initials: M
authors.lastname: Crossley
authors.forename: J J
authors.initials: JJ
authors.lastname: Bieker
authors.forename: S P
authors.initials: SP
authors.lastname: Klinken
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002455:Cell Division; D002461:Cell Line, Transformed; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D016373:DNA, Antisense; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004921:Erythropoietin / Q000494:pharmacology*; D005914:Globins / Q000096:biosynthesis / Q000235:genetics*; D006454:Hemoglobins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D051741:Kruppel-Like Transcription Factors; D004915:Leukemia, Erythroblastic, Acute; D051379:Mice; D009857:Oncogenes; D011994:Recombinant Proteins; D015398:Signal Transduction; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D016335:Zinc Fingers
chemical_list: D016373:DNA, Antisense; D004268:DNA-Binding Proteins; D006454:Hemoglobins; D051741:Kruppel-Like Transcription Factors; D011994:Recombinant Proteins; D014157:Transcription Factors; C080774:erythroid Kruppel-like factor; D004921:Erythropoietin; D005914:Globins