An erythroid specific enhancer upstream to the gene encodin the cell-type specific transcription factor GATA-1

Nicolis, Silvia K.; Bertini, C; Ronchi, Antonella; Crotta, Stefania; Lanfranco, Luisa; Moroni, Emanuela; Giglioni, Barbára; Ottolenghi, Sergio

doi:10.1093/nar/19.19.5285

Cited by 79 publications

(71 citation statements)

References 30 publications

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“…Initiator sequences, however, are located close to the transcription start sites of the IEb/c and IEd exons. By contrast, strong cis-acting motifs, such as the double GATA motif, the CP2-binding motif, and the CACCC motif, are located proximal to the IE exon (9,10,33). We speculate that these might contribute to the high-level transcription of the IE exon in erythroid cells.…”

Section: Discussionmentioning

confidence: 85%

“…the IT, IEb/c, and IEd, and their affiliated promoters are rarely used in these cell lineages (9,10,13). To further delineate the in vivo contribution of the IE exon to the maintenance of erythroid homeostasis, we conditionally deleted the IE exon in adulthood.…”

Section: Discussionmentioning

confidence: 99%

“…3 Of the first exons, the erythroid first exon (IE exon) is located 3.9 kb upstream of the second exon and is mainly utilized for transcription of the Gata1 gene in hematopoietic tissues (9,10). The distal testis first exon (IT exon) plays a role in Gata1 gene expression in mouse testis (11,12).…”

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confidence: 99%

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Loss of the Gata1 Gene IE Exon Leads to Variant Transcript Expression and the Production of a GATA1 Protein Lacking the N-terminal Domain

Kobayashi

Shimizu

Kikuchi

et al. 2010

Journal of Biological Chemistry

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GATA1 is essential for the differentiation of erythroid cells and megakaryocytes. The Gata1 gene is composed of multiple untranslated first exons and five common coding exons. The erythroid first exon (IE exon) is important for Gata1 gene expression in hematopoietic lineages. Because previous IE exon knockdown analyses resulted in embryonic lethality, less is understood about the contribution of the IE exon to adult hematopoiesis. Here, we achieved specific deletion of the floxed IE exon in adulthood using an inducible Cre expression system. In this conditional knock-out mouse line, the Gata1 mRNA level was significantly down-regulated in the megakaryocyte lineage, resulting in thrombocytopenia with a marked proliferation of megakaryocytes. By contrast, in the erythroid lineage, Gata1 mRNA was expressed abundantly utilizing alternative first exons. Especially, the IEb/c and newly identified IEd exons were transcribed at a level comparable with that of the IE exon in control mice. Surprisingly, in the IE-null mouse, these transcripts failed to produce full-length GATA1 protein, but instead yielded GATA1 lacking the N-terminal domain inefficiently. With low level expression of the short form of GATA1, IE-null mice showed severe anemia with skewed erythroid maturation. Notably, the hematological phenotypes of adult IE-null mice substantially differ from those observed in mice harboring conditional ablation of the entire Gata1 gene. The present study demonstrates that the IE exon is instrumental to adult erythropoiesis by regulating the proper level of transcription and selecting the correct transcription start site of the Gata1 gene.

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Section: Discussionmentioning

confidence: 85%

Section: Discussionmentioning

confidence: 99%

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Loss of the Gata1 Gene IE Exon Leads to Variant Transcript Expression and the Production of a GATA1 Protein Lacking the N-terminal Domain

Kobayashi

Shimizu

Kikuchi

et al. 2010

Journal of Biological Chemistry

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“…The remaining five exons, which encode the GATA-1 protein, are used in both Sertoli and erythroid mRNAs. Both the IE and IT proximal promoters were shown to contain duplicated GATA binding sites, and results from co-transfection assays suggested that GATA-1 gene expression may be regulated by GATA-1 itself or other GATA family members in both cell types (Hannon et al 1991;Nicolis et al 1991;Tsai et al 1991;Onodera et al 1997b). However, these transfection studies provided only limited physiological insight into the mechanism of GATA-1 gene regulation in vivo.…”

Section: An Upstream Activation Element Directs Primitive Haematopoiementioning

confidence: 99%

Upstream and downstream of erythroid transcription factor GATA‐1

et al. 1997

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All mature blood lineages in the peripheral circulation are derived from pluripotent haematopoietic stem cell. Progressive lineage-restriction of this stem cell is executed, in part, by the interplay and cross-talk between a host of lineage-restricted as well as ubiquitous transcription factors. To elucidate the regulatory mechanisms underlying the erythroid gene regulation, it is essential to understand how individual transcription factors contribute to the regulation of specific target genes, and how these erythroid transcription factor genes are regulated in turn. These key issues of mammalian development have been addressed by examining the activities controlling the prototype transcription factor, GATA-1. The transcriptional regulation of GATA-1 has been intensively investigated, thereby leading to the identification of its developmental stage-specific regulatory sequences. Loss-of-function mutant animals, combined with specific marking of the primitive and definitive erythroid lineages have also shed new insight into how GATA-1 activity is required in vivo at specific developmental stages. Procedures have also been developed for ascertaining whether or not the GATA-1 protein actually binds in vivo to regulatory GATA motifs in candidate target genes. Application of a similar multifaceted approach should enable investigators to examine the physiological roles that any transcription factor might play in vivo during the differentiation of any well defined cell lineage.

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“…The regulation of the expression of GATA-1 itself has been the subject of many investigations (7)(8)(9)(10)(11)(12)(13)(14)(15)(16). Constructs including the mouse GATA-1 promoter up to a DNase I-hypersensitive site lying at about Ϫ700 nts 1 (HS2) are expressed, at low efficiency, in adult hematopoietic cells in transgenic mice but not in yolk sac cells; however, constructs including an additional more upstream site (HS1) are much more efficient and are also active in primitive hematopoietic cells (12)(13)(14)(15)(16).…”

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confidence: 99%

Deletion of a Negatively Acting Sequence in a Chimeric GATA-1 Enhancer-Long Terminal Repeat Greatly Increases Retrovirally Mediated Erythroid Expression

Testa

Lotti

Cairns

et al. 2004

Journal of Biological Chemistry

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The locus control region of the ␤-globin gene cluster has been used previously to direct erythroid expression of globin genes from retroviral vectors for the purpose of gene therapy. Short erythroid regulatory elements represent a potentially valuable alternative to the locus control region. Among them, the GATA-1 enhancer HS2 was used to replace the retroviral enhancer within the 3 -long terminal repeat (LTR) of the retroviral vector SFCM, converting it into an erythroid-specific regulatory element. In this work, we have functionally studied an additional GATA-1 enhancer, HS1. HS1 participates in the transcriptional autoregulation of GATA-1 through an essential GATA-binding site that is footprinted in vivo. In this work we identified within HS1 a new in vivo footprinted region, and we showed that this sequence indeed binds a nuclear protein in vitro. Addition of HS1 to HS2 within the LTR of SFCM significantly improves the expression of a reporter gene. The deletion of the newly identified footprinted sequence in the retroviral construct further increases expression up to a level almost equal to that of the wild type retroviral LTR, without loss of erythroid specificity, suggesting that this sequence may act as a negative regulatory element. An improved vector backbone, M⌬N, allows even better expression from the new GATA cassette. These results suggest that substantial improvement of overall expression can be achieved by the combination of multiple changes in both regulatory elements and vectors.

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An erythroid specific enhancer upstream to the gene encodin the cell-type specific transcription factor GATA-1

Cited by 79 publications

References 30 publications

Loss of the Gata1 Gene IE Exon Leads to Variant Transcript Expression and the Production of a GATA1 Protein Lacking the N-terminal Domain

Loss of the Gata1 Gene IE Exon Leads to Variant Transcript Expression and the Production of a GATA1 Protein Lacking the N-terminal Domain

Upstream and downstream of erythroid transcription factor GATA‐1

Deletion of a Negatively Acting Sequence in a Chimeric GATA-1 Enhancer-Long Terminal Repeat Greatly Increases Retrovirally Mediated Erythroid Expression

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