NDLI: Characterization of a functional ZBP-89 binding site that mediates Gata1 gene expression during hematopoietic development.

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Characterization of a functional ZBP-89 binding site that mediates Gata1 gene expression during hematopoietic development.

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Characterization of a functional ZBP-89 binding site that mediates Gata1 gene expression during hematopoietic development.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Ohmori, Shin'ya Ishijima, Yasushi Ohneda, Kinuko Yamamoto, Masayuki Nakano, Mayu
Description	Author Affiliation: Ohneda K ( Department of Pharmacy, Faculty of Pharmacy, Takasaki University of Health and Welfare, Takasaki 370-0033, Japan. kohneda@takasaki-u.ac.jp)
Abstract	GATA-1 is a lineage-restricted transcription factor that plays essential roles in hematopoietic development. The Gata1 gene hematopoietic enhancer allowed Gata1 reporter expression in erythroid cells and megakaryocytes of transgenic mice. The Gata1 hematopoietic enhancer activity is strictly dependent on a GATA site located in the 5' region of the enhancer. However, the importance of the GC-rich region adjacent to the 3'-end of this GATA site has been also suggested. In this study, we show that this GC-rich region contains five contiguous deoxyguanosine residues (G(5) string) that are bound by multiple nuclear proteins. Interestingly, deletion of one deoxyguanosine residue from the G(5) string (G(4) mutant) specifically eliminates binding to ZBP-89, a Krüppel-like transcription factor, but not to Sp3 and other binding factors. We demonstrate that GATA-1 and ZBP-89 occupy chromatin regions of the Gata1 enhancer and physically associate in vitro through zinc finger domains. Gel mobility shift assays and DNA affinity precipitation assays suggest that binding of ZBP-89 to this region is reduced in the absence of GATA-1 binding to the G1HE. Luciferase reporter assays demonstrate that ZBP-89 activates the Gata1 enhancer depending on the G(5) string sequence. Finally, transgenic mouse studies reveal that the G(4) mutation significantly reduced the reporter activity of the Gata1 hematopoietic regulatory domain encompassing an 8.5-kbp region of the Gata1 gene. These data provide compelling evidence that the G(5) string is necessary for Gata1 gene expression in vivo and ZBP-89 is the functional trans-acting factor for this cis-acting region.
ISSN	00219258
e-ISSN	1083351X
Journal	Journal of Biological Chemistry
Issue Number	44
Volume Number	284
Language	English
Publisher	American Society for Biochemistry and Molecular Biology (United States)
Publisher Date	2009-10-30
Publisher Place	United States
Access Restriction	Open
Subject Keyword	DNA-Binding Proteins Metabolism Enhancer Elements, Genetic Genetics GATA1 Transcription Factor Hematopoiesis Transcription Factors Animals Binding Sites Chromatin Deoxyguanosine Erythroid Cells Biosynthesis Megakaryocytes Mice Mice, Transgenic Mutation Research Support, Non-U.S. Gov't Biochemistry Molecular Biology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology

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