TY - JOUR
T1 - The Gata1 5′ region harbors distinct cis-regulatory modules that direct gene activation in erythroid cells and gene inactivation in HSCs
AU - Takai, Jun
AU - Moriguchi, Takashi
AU - Suzuki, Mikiko
AU - Yu, Lei
AU - Ohneda, Kinuko
AU - Yamamoto, Masayuki
N1 - Funding Information:
We thank Dr Hozumi Motohashi and Ms Makiko Hayashi for their helpful discussion and assistance. This study was supported in part by the Grants-in-Aids for Scientific Research for the Scientific Research on Priority Areas (to M.Y.), by Scientific Research (to T.M. and M.Y.), by Exploratory Research (to M.Y.) from the Ministry of Education, Science, Sports and Culture, Naito foundation (to M.Y.), and by the Takeda Foundation (to T.M. and M.Y.). This study was also supported by the Global-COE program to conquest disease through network medicine. We also thank the Biomedical Research Core of Tohoku University Graduate School of Medicine for its technical support.
Funding Information:
This study was supported in part by the Grants-in-Aids for Scientific Research for the Scientific Research on Priority Areas (to M.Y.), by Scientific Research (to T.M. and M.Y.), by Exploratory Research (to M.Y.) from the Ministry of Education, Science, Sports and Culture, Naito foundation (to M.Y.), and by the Takeda Foundation (to T.M. and M.Y.). This study was also supported by the Global-COE program to conquest disease through network medicine. We also thank the Biomedical Research Core of Tohoku University Graduate School of Medicine for its technical support.
PY - 2013
Y1 - 2013
N2 - GATA1 is a master regulator of hematopoietic differentiation, but Gata1 expression is inactivated in hematopoietic stem cells (HSCs). Using a bacterial artificial chromosome containing the Gata1 gene modified with green fluorescent protein (GFP) reporter, we explored the function of the 3.7-kb Gata1 upstream region (GdC region) that harbors 3 core cis-elements: Gata1 hematopoietic enhancer, double GATA-motif, and CACCC-motif. Transgenic GFP expression directed by the Gata1-BAC faithfully recapitulated the endogenous Gata1 expression pattern. However, deletion of the GdC-region eliminated reporter expression in all hematopoietic cells. To test whether the combination of the core cis-elements represents the regulatory function of the GdC-region, we replaced the region with a 659-bp minigene that linked the three cis-elements (MG-GFP). The GFP reporter expression directed by the MG-GFP BAC fully recapitulated the erythroid-megakaryocytic Gata1 expression. However, the GFP expression was aberrantly increased in the HSCs and was associated with decreases in DNA methylation and abundant GATA2 binding to the transgenic MG-GFP allele. The 3.2-kb sequences interspaced between the Gata1 hematopoietic enhancer and the double GATA-motif were able to recruit DNA methyltransferase 1, thereby exerting a cis-repressive function in the HSC-like cell line. These results indicate that the 3.2-kb interspacing sequences inactivate Gata1 by maintaining DNA-methylation in the HSCs.
AB - GATA1 is a master regulator of hematopoietic differentiation, but Gata1 expression is inactivated in hematopoietic stem cells (HSCs). Using a bacterial artificial chromosome containing the Gata1 gene modified with green fluorescent protein (GFP) reporter, we explored the function of the 3.7-kb Gata1 upstream region (GdC region) that harbors 3 core cis-elements: Gata1 hematopoietic enhancer, double GATA-motif, and CACCC-motif. Transgenic GFP expression directed by the Gata1-BAC faithfully recapitulated the endogenous Gata1 expression pattern. However, deletion of the GdC-region eliminated reporter expression in all hematopoietic cells. To test whether the combination of the core cis-elements represents the regulatory function of the GdC-region, we replaced the region with a 659-bp minigene that linked the three cis-elements (MG-GFP). The GFP reporter expression directed by the MG-GFP BAC fully recapitulated the erythroid-megakaryocytic Gata1 expression. However, the GFP expression was aberrantly increased in the HSCs and was associated with decreases in DNA methylation and abundant GATA2 binding to the transgenic MG-GFP allele. The 3.2-kb sequences interspaced between the Gata1 hematopoietic enhancer and the double GATA-motif were able to recruit DNA methyltransferase 1, thereby exerting a cis-repressive function in the HSC-like cell line. These results indicate that the 3.2-kb interspacing sequences inactivate Gata1 by maintaining DNA-methylation in the HSCs.
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U2 - 10.1182/blood-2013-01-476911
DO - 10.1182/blood-2013-01-476911
M3 - Article
C2 - 24021675
AN - SCOPUS:84888218999
VL - 122
SP - 3450
EP - 3460
JO - Blood
JF - Blood
SN - 0006-4971
IS - 20
ER -