Alternate Promoters and Developmental Modulation of Expression of the Chicken GATA-2 Gene in Hematopoietic Progenitor Cells

Nony, Pascale; Hannon, Robert; Gould, Hannah; Felsenfeld, Gary

doi:10.1074/jbc.273.49.32910

Cited by 16 publications

(15 citation statements)

References 48 publications

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“…We find that by contrast to chicken GATA-6, which is transcribed from a single promoter (9), the mammalian genes both contain two alternative noncoding upstream exons, transcribed from two distinct promoters, as reported previously for some other members of the GATA family (2,10,11,12). By a combination of RNase protection analysis, exon-specific in situ hybridization, and RT-PCR, we have not detected any difference in the spatial expression patterns of the two GATA-6 transcript isoforms.…”

supporting

confidence: 83%

“…Similarly, an upstream promoter within the mouse GATA-2 gene is active only in hematopoietic cells, while a downstream region directs GATA-2 transcription in all cells in which it is expressed (10). Two distinct transcriptional start sites have also been reported for the chicken GATA-2 gene, although the tissue distribution of promoter usage differs from the mouse (9,11). Most notably, the predominant chicken GATA-2 transcript within erythroid cells derives from the proximal promoter, demonstrating evolutionary diversity in the regulation of the GATA-2 gene between avian and mammalian species.…”

Section: The 5ј-utr Does Not Affect the Rate Or Site Of Translationalmentioning

confidence: 98%

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The Human and Mouse GATA-6 Genes Utilize Two Promoters and Two Initiation Codons

Brewer¹,

Gove²,

Davies³

et al. 1999

Journal of Biological Chemistry

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GATA-6 has been implicated in the regulation of myocardial differentiation during cardiogenesis. To determine how its expression is controlled, we have characterized the human and mouse genes. We have mapped their transcriptional start sites and demonstrate that two alternative promoters and 5 noncoding exons are utilized. Both transcript isoforms are expressed in the same tissue-specific and developmental stage-specific pattern, and their ratio appears similar wherever examined. The more upstream noncoding exon showed a substantial degree of homology between the two mammalian species, suggesting a conserved regulatory function. Moreover, in transfection assays we show that elements within this exon act to promote its transcription. Positive regulatory elements that effect transcription from the more downstream exon were not apparent in this assay, revealing a regulatory distinction between the two promoters. We also demonstrate alternative initiator codon usage in both the human and mouse GATA-6 genes. Both isoforms of the protein are synthesized in vitro regardless of which 5 noncoding exon is present in the RNA, although the larger protein has greater transcriptional activation potential in transfection assays. Thus, GATA-6 function in the cell is controlled by a complex interplay of transcriptional and translational regulation.

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supporting

confidence: 83%

Section: The 5ј-utr Does Not Affect the Rate Or Site Of Translationalmentioning

confidence: 98%

The Human and Mouse GATA-6 Genes Utilize Two Promoters and Two Initiation Codons

Brewer¹,

Gove²,

Davies³

et al. 1999

Journal of Biological Chemistry

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“…Interestingly, this intron contains a sequence motif homologous to a regular TATA box (TATAAA) near its 3¢ border. This sequence element could be used as an internal alternative promoter, a situation that has been previously identified in other genes containing an intron within their 5¢ UTR [33]. Alternative splicing and/or use of alternate promoters could contribute to explain previously reported size differences in MGP mRNAs [34,35].…”

Section: Discussionmentioning

confidence: 77%

Molecular cloning of the Matrix Gla Protein gene from Xenopus laevis. Functional analysis of the promoter identifies a calcium sensitive region required for basal activity

et al. 2002

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To analyze the regulation of Matrix Gla Protein (MGP) gene expression in Xenopus laevis, we cloned the xMGP gene and its 5¢ region, determined their molecular organization, and characterized the transcriptional properties of the core promoter. The Xenopus MGP (xMGP) gene is organized into five exons, one more as its mammalian counterparts. The first two exons in the Xenopus gene encode the DNA sequence that corresponds to the first exon in mammals whereas the last three exons show homologous organization in the Xenopus MGP gene and in the mammalian orthologs. We characterized the transcriptional regulation of the xMGP gene in transient transfections using Xenopus A6 cells. In our assay system the identified promoter was shown to be transcriptionally active, resulting in a 12-fold induction of reporter gene expression. Deletional analysis of the 5¢ end of the xMGP promoter revealed a minimal activating element in the sequence from )70 to )36 bp. Synthetic reporter constructs containing three copies of the defined regulatory element delivered 400-fold superactivation, demonstrating its potential for the recruitment of transcriptional activators. In gel mobility shift assays we demonstrate binding of X. laevis nuclear factors to an extended regulatory element from )180 to )36, the specificity of the interaction was proven in competition experiments using different fragments of the xMGP promoter. By this approach the major site of factor binding was demonstrated to be included in the minimal activating promoter fragment from )70 to )36 bp. In addition, in transient transfection experiments we could show that this element mediates calcium dependent transcription and increasing concentrations of extracellular calcium lead to a significant dose dependent activation of reporter gene expression.

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“…The gene-distal first exon (IS) is specifically expressed in hematopoietic and neural cells, whereas the geneproximal first exon (IG) is transcribed in almost all Gata2-expressing cells (29). Differential utilization of two distinct first exons has also been demonstrated for the human and chicken Gata2 genes (37,44). We also reported that the murine Gata2 gene contains a number of hematopoietic regulatory elements scattered over more than 250 kbp of the locus, since a 250-kbp yeast artificial chromosome transgene was capable of rescuing the hematopoietic lethal deficiency in Gata2 null mutant embryos (66).…”

mentioning

confidence: 84%

GATA Motifs Regulate Early Hematopoietic Lineage-Specific Expression of the Gata2 Gene

Kobayashi-Osaki

Onodera

Suzuki

et al. 2005

Molecular and Cellular Biology

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Transcription factor GATA-2 is essential for definitive hematopoiesis, which developmentally emerges from the para-aortic splanchnopleura (P-Sp). The expression of a green fluorescent protein (GFP) reporter placed under the control of a 3.1-kbp Gata2 gene regulatory domain 5 to the distal first exon (IS) mirrored that of the endogenous Gata2 gene within the P-Sp and yolk sac (YS) blood islands of embryonic day (E) 9.5 murine embryos. The P-Sp-and YS-derived GFP ؉ fraction of flow-sorted cells dissociated from E9.5 transgenic embryos contained far more CD34؉ /c-Kit ؉ cells than the GFP ؊ fraction did. When cultured in vitro, the P-Sp GFP ؉ cells generated both immature hematopoietic and endothelial cell clusters. Detailed transgenic mouse reporter expression analyses demonstrate that five GATA motifs within the 3.1-kbp Gata2 early hematopoietic regulatory domain (G2-EHRD) were essential for GFP expression within the dorsal aortic wall, where hemangioblasts, the earliest precursors possessing both hematopoietic and vascular developmental potential, are thought to reside. These results thus show that the Gata2 gene IS promoter is regulated by a GATA factor(s) and selectively marks putative hematopoietic/endothelial precursor cells within the P-Sp.

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Alternate Promoters and Developmental Modulation of Expression of the Chicken GATA-2 Gene in Hematopoietic Progenitor Cells

Cited by 16 publications

References 48 publications

The Human and Mouse GATA-6 Genes Utilize Two Promoters and Two Initiation Codons

The Human and Mouse GATA-6 Genes Utilize Two Promoters and Two Initiation Codons

Molecular cloning of the Matrix Gla Protein gene from Xenopus laevis. Functional analysis of the promoter identifies a calcium sensitive region required for basal activity

GATA Motifs Regulate Early Hematopoietic Lineage-Specific Expression of the Gata2 Gene

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