title: Establishment of erythroleukemic GAK14 cells and characterization of GATA1 N-terminal domain.
abstract: GATA1 is a transcription factor essential for erythropoiesis and megakaryopoiesis. It has been found that Gata1 gene knockdown heterozygous female (Gata1(G1.05/+)) mice spontaneously develop erythroblastic leukemias. In this study, we have generated a novel Gata1 knockdown erythroblastic cell line, designated GAK14, from the leukemia cells in the Gata1(G1.05/+) mice. Although GAK14 cells maintain immature phenotype on OP9 stromal cells in the presence of erythropoietin and stem cell factor, the cells produce Gr-1-, Mac1-, B220-, CD3e- or CD49b-positive hematopoietic cells when co-cultured with DAS104-8 feeder cells. However, GAK14 cells did not produce erythroid and megakaryocytic lineages, perhaps due to the absence of GATA1. Indeed, GAK14 cells became capable of differentiating into mature erythroid cells when complemented with full-length GATA1 and co-cultured with fetal liver-derived FLS5 stromal cells. This differentiation potential was impaired when GATA1 lacking the N-terminal domain was complemented. The N-terminal domain is known to contribute to the pathogenesis of transient abnormal myelopoiesis and acute megakaryoblastic leukemia related to Down syndrome. These results thus showed that GAK14 cells will serve as a powerful tool for dissecting domain function of GATA1 and that the GATA1 N-terminal domain is essential for the erythroid differentiation of GAK14 cells.
authors.affiliation: Graduate School of Comprehensive Human Sciences, Center for Tsukuba Advanced Research Alliance, University of Tsukuba, Tsukuba, 305-8577, Japan; Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, Sendai, 980-8575, Japan.
authors.forename: Harumi Y
authors.initials: HY
authors.lastname: Mukai
authors.forename: Mikiko
authors.initials: M
authors.lastname: Suzuki
authors.forename: Masumi
authors.initials: M
authors.lastname: Nagano
authors.forename: Shin'ya
authors.initials: S
authors.lastname: Ohmori
authors.forename: Akihito
authors.initials: A
authors.lastname: Otsuki
authors.forename: Kouhei
authors.initials: K
authors.lastname: Tsuchida
authors.forename: Takashi
authors.initials: T
authors.lastname: Moriguchi
authors.forename: Kinuko
authors.initials: K
authors.lastname: Ohneda
authors.forename: Ritsuko
authors.initials: R
authors.lastname: Shimizu
authors.forename: Osamu
authors.initials: O
authors.lastname: Ohneda
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yamamoto
mesh_terms: D000596:Amino Acids / Q000235:genetics*; D000818:Animals; D045744:Cell Line, Tumor; D019070:Cell Lineage; D018920:Coculture Techniques; D015672:Erythroid Precursor Cells / Q000378:metabolism / Q000502:physiology*; D004920:Erythropoiesis; D005260:Female; D050982:GATA1 Transcription Factor / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D004915:Leukemia, Erythroblastic, Acute; D055016:Megakaryocyte Progenitor Cells / Q000502:physiology; D051379:Mice; D038042:Myelopoiesis; D017434:Protein Structure, Tertiary
chemical_list: D000596:Amino Acids; D050982:GATA1 Transcription Factor; C494689:Gata1 protein, mouse