A GATA Box in the GATA-1 Gene Hematopoietic Enhancer Is a Critical Element in the Network of GATA Factors and Sites That Regulate This Gene

ABSTRACT A region located at kbp −3.9 to −2.6 5′ to the first hematopoietic exon of the GATA-1 gene is necessary to recapitulate gene expression in both the primitive and definitive erythroid lineages. In transfection analyses, this region activated reporter gene expression from an artificial promoter in a position- and orientation-independent manner, indicating that the region functions as the GATA-1 gene hematopoietic enhancer (G1HE). However, when analyzed in transgenic embryos in vivo, G1HE activity was orientation dependent and also required the presence of the endogenousGATA-1 gene hematopoietic promoter. To define the boundaries of G1HE, a series of deletion constructs were prepared and tested in transfection and transgenic mice analyses. We show that G1HE contains a 149-bp core region which is critical for GATA-1gene expression in both primitive and definitive erythroid cells but that expression in megakaryocytes requires the core plus additional sequences from G1HE. This core region contains one GATA, one GAT, and two E boxes. Mutational analyses revealed that only the GATA box is critical for gene-regulatory activity. Importantly, G1HE was active in SCL−/− embryos. These results thus demonstrate the presence of a critical network of GATA factors and GATA binding sites that controls the expression of this gene.

[1]  J. D. Engel,et al.  The mouse GATA-2 gene is expressed in the para-aortic splanchnopleura and aorta-gonads and mesonephros region. , 1999, Blood.

[2]  O. Nakajima,et al.  Role of GATA-1 in proliferation and differentiation of definitive erythroid and megakaryocytic cells in vivo. , 1998, Blood.

[3]  L. Madisen,et al.  Identification of Bach2 as a B‐cell‐specific partner for small Maf proteins that negatively regulate the immunoglobulin heavy chain gene 3′ enhancer , 1998, The EMBO journal.

[4]  C. Begley,et al.  Absence of yolk sac hematopoiesis from mice with a targeted disruption of the scl gene. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[5]  S. Orkin,et al.  Functional analysis and in vivo footprinting implicate the erythroid transcription factor GATA-1 as a positive regulator of its own promoter. , 1991, Genes & development.

[6]  J. T. Kadonaga,et al.  *To whom correspondence should be addressed. E- , 2022 .

[7]  J. D. Engel,et al.  Activity and tissue-specific expression of the transcription factor NF-E1 multigene family. , 1990, Genes & development.

[8]  J. D. Engel,et al.  Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain. , 1999, Genes & development.

[9]  J. D. Engel,et al.  Dynamics of GATA transcription factor expression during erythroid differentiation. , 1993, Blood.

[10]  S. Orkin,et al.  A lineage‐selective knockout establishes the critical role of transcription factor GATA‐1 in megakaryocyte growth and platelet development , 1997, The EMBO journal.

[11]  Satoru Takahashi,et al.  Differential roles of GATA‐1 and GATA‐2 in growth and differentiation of mast cells , 1998, Genes to cells : devoted to molecular & cellular mechanisms.

[12]  J. D. Engel,et al.  GATA-1 transcription is controlled by distinct regulatory mechanisms during primitive and definitive erythropoiesis. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[13]  P. Chambon,et al.  The SV40 72 base repair repeat has a striking effect on gene expression both in SV40 and other chimeric recombinants. , 1981, Nucleic acids research.

[14]  G. Keller,et al.  Rescue of erythroid development in gene targeted GATA–1− mouse embryonic stem cells , 1992, Nature Genetics.

[15]  Stuart H. Orkin,et al.  Expression of an erythroid transcription factor in megakaryocytic and mast cell lineages , 1990, Nature.

[16]  J. D. Engel,et al.  Conserved structure, regulatory elements, and transcriptional regulation from the GATA-1 gene testis promoter. , 1997, Journal of biochemistry.

[17]  P. Ricciardi-Castagnoli,et al.  Molecular heterogeneity of regulatory elements of the mouse GATA-1 gene. , 1997, Genes and function.

[18]  S. Orkin,et al.  Development of hematopoietic cells lacking transcription factor GATA-1. , 1995, Development.

[19]  S. Orkin,et al.  Unsuspected role for the T-cell leukemia protein SCL/tal-1 in vascular development. , 1998, Genes & development.

[20]  A. Gronenborn,et al.  A palindromic regulatory site within vertebrate GATA-1 promoters requires both zinc fingers of the GATA-1 DNA-binding domain for high-affinity interaction , 1996, Molecular and cellular biology.

[21]  Stuart H. Orkin,et al.  An early haematopoietic defect in mice lacking the transcription factor GATA-2 , 1994, Nature.

[22]  S. Nicolis,et al.  An erythroid specific enhancer upstream to the gene encoding the cell-type specific transcription factor GATA-1. , 1991, Nucleic Acids Research.

[23]  W. Vainchenker,et al.  Megakaryocytic and erythrocytic lineages share specific transcription factors , 1990, Nature.

[24]  L. Zon,et al.  Expression of mRNA for the GATA-binding proteins in human eosinophils and basophils: potential role in gene transcription. , 1993, Blood.

[25]  J. D. Engel,et al.  Transcription factor GATA-2 is expressed in erythroid, early myeloid, and CD34+ human leukemia-derived cell lines. , 1994, Blood.

[26]  Masayuki Yamamoto,et al.  Upstream and downstream of erythroid transcription factor GATA‐1 , 1997, Genes to cells : devoted to molecular & cellular mechanisms.

[27]  G. Felsenfeld,et al.  Structure and promoter activity of the gene for the erythroid transcription factor GATA-1. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[28]  J. D. Engel,et al.  Developmental stage- and spermatogenic cycle-specific expression of transcription factor GATA-1 in mouse Sertoli cells. , 1994, Development.

[29]  M. Yamamoto,et al.  GATA-1 regulates growth and differentiation of definitive erythroid lineage cells during in vitro ES cell differentiation. , 1998, Blood.

[30]  J. D. Engel,et al.  Erythroid transcription factor GATA-1 is abundantly transcribed in mouse testis , 1993, Nature.

[31]  M. Groudine,et al.  Transcriptional regulation of hemoglobin switching in chicken embryos , 1981, Molecular and cellular biology.

[32]  N. Hayashi,et al.  Arrest in Primitive Erythroid Cell Development Caused by Promoter-specific Disruption of the GATA-1 Gene* , 1997, The Journal of Biological Chemistry.

[33]  T. Rabbitts,et al.  The LIM‐only protein Lmo2 is a bridging molecule assembling an erythroid, DNA‐binding complex which includes the TAL1, E47, GATA‐1 and Ldb1/NLI proteins , 1997, The EMBO journal.

[34]  S. Orkin,et al.  Absence of blood formation in mice lacking the T-cell leukaemia oncoprotein tal-1/SCL , 1995, Nature.