Transcription of the SCL gene in erythroid and CD34 positive primitive myeloid cells is controlled by a complex network of lineage-restricted chromatin-dependent and chromatin-independent regulatory elements

Göttgens, Berthold; McLaughlin, Fiona; Bockamp, Ernesto; Fordham, Jeremy; Begley, C. Glenn; Kosmopoulos, K; Elefanty, Andrew G.; Green, Anthony

doi:10.1038/sj.onc.1201426

Cited by 56 publications

(78 citation statements)

References 54 publications

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“…The fact that the 5,245-bp fragment was not prone to β-galactosidase-mediated transgene inactivation may be due to the presence of one or more additional regulatory elements in this bigger fragment. A strong candidate for such an element is the +18 region, which adopts an open chromatin configuration in progenitor cells [15] and can enhance activity of the +19 core enhancer in transfection assays (L.S. and B.G., unpublished data).…”

Section: Discussionmentioning

confidence: 99%

“…So far, these studies have resulted in the identification of five independent enhancers, each of which targets expression to a specific subdomain of the normal SCL expression pattern [14][15][16][17][18]. Of particular note, a 3' element (the SCL +18/19 enhancer) was found to direct expression to embryonic endothelium and to most adult and embryonic hematopoietic progenitors and long-term repopulating stem cells [18,19].…”

Section: Introductionmentioning

confidence: 99%

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Transgenic Analysis of the Stem Cell Leukemia +19 Stem Cell Enhancer in Adult and Embryonic Hematopoietic and Endothelial Cells

et al. 2005

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Appropriate transcriptional regulation is critical for the biological functions of many key regulatory genes, including the stem cell leukemia (SCL) gene. As part of a systematic dissection of SCL transcriptional regulation, we have previously identified a 5,245-bp SCL +18/19 enhancer that targeted embryonic endothelium together with embryonic and adult hematopoietic progenitors and stem cells (HSCs). This enhancer is proving to be a powerful tool for manipulating hematopoietic progenitors and stem cells, but the design and interpretation of such transgenic studies require a detailed understanding of enhancer activity in vivo. In this study, we demonstrate that the +18/19 enhancer is active in mast cells, megakaryocytes, and adult endothelium. A 644-bp +19 core enhancer exhibited similar temporal and spatial activity to the 5,245-bp +18/19 fragment both during development and in adult mice. Unlike the +18/19 enhancer, the +19 core enhancer was only active in adult mice when linked to the eukaryotic reporter gene human placental alkaline phosphatase. Activity of a single core enhancer in HSCs, endothelium, mast cells, and megakaryocytes suggests possible overlaps in their respective transcriptional programs. Moreover, activity in a proportion of thymocytes and other SCL-negative cell types suggests the existence of a silencer elsewhere in the SCL locus.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Transgenic Analysis of the Stem Cell Leukemia +19 Stem Cell Enhancer in Adult and Embryonic Hematopoietic and Endothelial Cells

et al. 2005

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“…1A). This strategy ensured that SCL regulatory regions upstream of exon III and downstream of exon VI were left intact (18). Correctly targeted ES cell clones were identified by Southern hybridization of DNA using the 3Ј flanking probe A.…”

Section: Generation Of Sclmentioning

confidence: 99%

Characterization of hematopoietic progenitor cells that express the transcription factor SCL, using a lacZ “knock-in” strategy

Elefanty

Begley

Metcalf

et al. 1998

Proc. Natl. Acad. Sci. U.S.A.

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Gene targeting experiments have demonstrated that the transcription factor SCL is essential for primitive and definitive hematopoiesis in the mouse. To study the functional properties of hematopoietic cells expressing SCL, we have generated mutant mice (SCL lacZ͞w ) in which the Escherichia coli lacZ reporter gene has been ''knocked in'' to the SCL locus, thereby linking ␤-galactosidase expression to transcription from the SCL promoter. Bone marrow cells from heterozygous SCL lacZ͞w mice were sorted into fractions expressing high, intermediate and low levels of ␤-galactosidase (designated lacZ high , lacZ int , and lacZ neg ). Cells that were lacZ high or lacZ int were enriched for day 12 spleen colonyforming units and myeloid and erythroid colony-forming cells (CFCs). These fractions included >99% of the erythroid and >90% of the myeloid CFCs. Culture of sorted bone marrow populations on stromal cells secreting interleukin-7 or in fetal thymic organ cultures showed that B and T lymphoid progenitors were also present in the lacZ high and lacZ int fractions. These data provide a functional correlation between SCL expression and colony-forming ability in immature hematopoietic cells. Our data also suggested that expression of SCL was transient and confined to hematopoietic stem and͞or progenitor cells, because the differentiated progeny of most lineages (except the erythroid) were ␤-galactosidase-negative.

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“…Both human and mouse SCL are transcribed from two lineage-specific promoters (20)(21)(22)(23)(24). In addition, a panel of DNase I hypersensitive sites associated with enhancer or silencer activity have been identified within the murine SCL locus (25,26). Studies using transgenic mice have subsequently defined a 30-kb region containing five distinct murine enhancers that are capable of directing reporter gene expression to hemopoietic progenitor cells, endothelium, and specific regions of the brain and spinal cord (6,27,28).…”

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confidence: 99%

Regulation of the stem cell leukemia ( SCL ) gene: A tale of two fishes

Barton

Göttgens

Gering

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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The stem cell leukemia (SCL) gene encodes a tissue-specific basic helix-loop-helix (bHLH) protein with a pivotal role in hemopoiesis and vasculogenesis. Several enhancers have been identified within the murine SCL locus that direct reporter gene expression to subdomains of the normal SCL expression pattern, and long-range sequence comparisons of the human and murine SCL loci have identified additional candidate enhancers. To facilitate the characterization of regulatory elements, we have sequenced and analyzed 33 kb of the SCL genomic locus from the pufferfish Fugu rubripes, a species with a highly compact genome. Although the pattern of SCL expression is highly conserved from mammals to teleost fish, the genes flanking pufferfish SCL were unrelated to those known to flank both avian and mammalian SCL genes. These data suggest that SCL regulatory elements are confined to the region between the upstream and downstream flanking genes, a region of 65 kb in human and 8.5 kb in pufferfish. Consistent with this hypothesis, the entire 33-kb pufferfish SCL locus directed appropriate expression to hemopoietic and neural tissue in transgenic zebrafish embryos, as did a 10.4-kb fragment containing the SCL gene and extending to the 5 and 3 flanking genes. These results demonstrate the power of combining the compact genome of the pufferfish with the advantages that zebrafish provide for studies of gene regulation during development. Furthermore, the pufferfish SCL locus provides a powerful tool for the manipulation of hemopoiesis and vasculogenesis in vivo.T he stem cell leukemia (SCL) gene (also known as Tal-1) encodes a basic helix-loop-helix (bHLH) transcription factor that is a critical regulator of both hemopoiesis and vasculogenesis (1). SCL is normally expressed in hemopoietic stem cells, in committed cells of the erythroid, myeloid, and megakaryocytic lineages, in endothelium, and within specific regions of the central nervous system, a pattern of expression that is highly conserved across vertebrate species from mammals to teleost fish (2-6).SCL is essential for the development of all hemopoietic lineages (7,8). A role in early progenitors is also suggested by the observation that expression of anti-sense SCL suppressed the proliferation, cell cycle progression, and self renewal of a multipotent hemopoietic cell line (9). Distinct functions following lineage commitment are implied by the modulation of SCL mRNA and protein levels during erythroid and myeloid differentiation (10-14). Moreover, enforced SCL expression enhanced erythroid differentiation of hemopoietic cell lines (14, 15) and increased erythroid and megakaryocytic differentiation of normal CD34-positive progenitors (16,17).SCL also plays a crucial role in the formation of hemangioblasts, bipotent progenitors of both blood and endothelium. Ectopic expression of SCL mRNA in zebrafish embryos specifies hemangioblast formation from early mesoderm and results in excessive production of blood and endothelial progenitors (3). In keeping with this, SCL can parti...

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Transcription of the SCL gene in erythroid and CD34 positive primitive myeloid cells is controlled by a complex network of lineage-restricted chromatin-dependent and chromatin-independent regulatory elements

Cited by 56 publications

References 54 publications

Transgenic Analysis of the Stem Cell Leukemia +19 Stem Cell Enhancer in Adult and Embryonic Hematopoietic and Endothelial Cells

Transgenic Analysis of the Stem Cell Leukemia +19 Stem Cell Enhancer in Adult and Embryonic Hematopoietic and Endothelial Cells

Characterization of hematopoietic progenitor cells that express the transcription factor SCL, using a lacZ “knock-in” strategy

Regulation of the stem cell leukemia ( SCL ) gene: A tale of two fishes

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