title: GATA2 is critical for the maintenance of cellular identity in differentiated mast cells derived from mouse bone marrow.
abstract: GATA2 plays a crucial role for the mast cell fate decision. We herein demonstrate that GATA2 is also required for the maintenance of the cellular identity in committed mast cells derived from mouse bone marrow (BMMCs). The deletion of the GATA2 DNA binding domain (GATA2ΔCF) in BMMCs resulted in a loss of the mast cell phenotype and an increase in the number of CD11b- and/or Ly6G/C-positive cells. These cells showed the ability to differentiate into macrophage- and neutrophil-like cells but not into eosinophils. Although the mRNA levels of basophil-specific genes were elevated, CD49b, a representative basophil marker, never appeared on these cells. GATA2 ablation led to a significant upregulation of C/EBPα, and forced expression of C/EBPα in wild-type BMMCs phenocopied the GATA2ΔCF cells. Interestingly, simultaneous deletion of the Gata2 and Cebpa genes in BMMCs restored the aberrant increases of CD11b and Ly6G/C while retaining the reduced c-Kit expression. Chromatin immunoprecipitation assays indicated that GATA2 directly binds to the +37-kb region of the Cebpa gene and thereby inhibits the RUNX1 and PU.1 binding to the neighboring region. Upregulation of C/EBPα following the loss of GATA2 was not observed in cultured mast cells derived from peritoneal fluid, whereas the repression of c-Kit and other mast cell-specific genes were observed in these cells. Collectively, these results indicate that GATA2 maintains cellular identity by preventing Cebpa gene activation in a subpopulation of mast cells, whereas it plays a fundamental role as a positive regulator of mast cell-specific genes throughout development of this cell lineage.
authors: Department of Pharmacy, Faculty of Pharmacy, Takasaki University of Health and Welfare, Takasaki, Japan;
authors: Shin'ya
authors: S
authors: Ohmori
authors: Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, Sendai, Japan; and.
authors: Takashi
authors: T
authors: Moriguchi
authors: Department of Pharmacy, Faculty of Pharmacy, Takasaki University of Health and Welfare, Takasaki, Japan;
authors: Yuki
authors: Y
authors: Noguchi
authors: Department of Pharmacy, Faculty of Pharmacy, Takasaki University of Health and Welfare, Takasaki, Japan;
authors: Muneharu
authors: M
authors: Ikeda
authors: Department of Pharmacy, Faculty of Pharmacy, Takasaki University of Health and Welfare, Takasaki, Japan;
authors: Kota
authors: K
authors: Kobayashi
authors: Department of Pharmacy, Faculty of Pharmacy, Takasaki University of Health and Welfare, Takasaki, Japan;
authors: Nazuki
authors: N
authors: Tomaru
authors: Department of Pharmacy, Faculty of Pharmacy, Takasaki University of Health and Welfare, Takasaki, Japan;
authors: Yasushi
authors: Y
authors: Ishijima
authors: Graduate School of Comprehensive Human Sciences, Laboratory of Regenerative Medicine and Stem Cell Biology, University of Tsukuba, Tsukuba, Japan.
authors: Osamu
authors: O
authors: Ohneda
authors: Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, Sendai, Japan; and.
authors: Masayuki
authors: M
authors: Yamamoto
authors: Department of Pharmacy, Faculty of Pharmacy, Takasaki University of Health and Welfare, Takasaki, Japan;
authors: Kinuko
authors: K
authors: Ohneda
mesh_terms: D000818:Animals; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000166:cytology*; D054337:Cell Dedifferentiation / Q000276:immunology*; D002454:Cell Differentiation / Q000276:immunology; D047369:Chromatin Immunoprecipitation; D005434:Flow Cytometry; D050989:GATA2 Transcription Factor / Q000276:immunology / Q000378:metabolism*; D008407:Mast Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology*
chemical_list: D050989:GATA2 Transcription Factor; C494712:Gata2 protein, mouse