Roles of fetalGγ-globin promoter elements and the adult β-globin 3′ enhancer in the stage-specific expression of globin genes

Carlos Perez-Stable, Frank Costantini

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The human fetal Gγ-globin and adult β-globin genes are expressed in a tissue- and developmental stage-specific pattern in transgenic mice: the Gγ gene in embryonic cells and the β gene in fetal and adult erythroid cells. Several of the cis-acting DNA sequences thought to be responsible for these patterns of expression are located 5′ to the Gγ-globin gene and 3′ to the β-globin gene. To further define the locations and functional roles of these elements, we examined the effects of 5′ truncations on the expression of the Gγ-globin gene, as well as the ability of Gγ-globin upstream sequences to alter the developmental regulation of a β-globin gene. We found that sequences between -201 and -136 are essential for expression of the Gγ-globin gene, whereas those upstream of -201 have little effect on the level or tissue or stage specificity of Gγ-globin expression. The Gγ-globin upstream sequences from -201 to -136 were, furthermore, capable of activating a linked β-globin gene in embryonic blood cells; however, a Gγ-globin fragment from -383 to -206 was similarly active in this assay, and the complete fragment from -383 to -136 was considerably more active than either of the smaller fragments, suggesting the presence of multiple cis-acting elements for embryonic blood cells. Our data also suggested the possibility of a negative regulatory element between -201 and -136. These results are discussed in relation to several DNA elements in the Gγ-globin upstream region, which have been shown to bind nuclear factors in erythroid cells. Finally, we observed that removal of the β-globin 3′-flanking sequences, including the 3′ enhancer, from the Gγ-globin upstream-β-globin hybrid gene resulted in a 25-fold reduction in expression in embryonic blood cells. This suggests that the β-globin 3′ enhancer is potentially active at the embryonic stage and thus cannot be solely responsible for the fetal or adult specificity of the β-globin gene.

Original languageEnglish
Pages (from-to)1116-1125
Number of pages10
JournalMolecular and Cellular Biology
Volume10
Issue number3
StatePublished - Dec 1 1990
Externally publishedYes

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Globins
Gene Expression
Genes
Blood Cells
Erythroid Cells
3' Flanking Region

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Roles of fetalGγ-globin promoter elements and the adult β-globin 3′ enhancer in the stage-specific expression of globin genes. / Perez-Stable, Carlos; Costantini, Frank.

In: Molecular and Cellular Biology, Vol. 10, No. 3, 01.12.1990, p. 1116-1125.

Research output: Contribution to journalArticle

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abstract = "The human fetal Gγ-globin and adult β-globin genes are expressed in a tissue- and developmental stage-specific pattern in transgenic mice: the Gγ gene in embryonic cells and the β gene in fetal and adult erythroid cells. Several of the cis-acting DNA sequences thought to be responsible for these patterns of expression are located 5′ to the Gγ-globin gene and 3′ to the β-globin gene. To further define the locations and functional roles of these elements, we examined the effects of 5′ truncations on the expression of the Gγ-globin gene, as well as the ability of Gγ-globin upstream sequences to alter the developmental regulation of a β-globin gene. We found that sequences between -201 and -136 are essential for expression of the Gγ-globin gene, whereas those upstream of -201 have little effect on the level or tissue or stage specificity of Gγ-globin expression. The Gγ-globin upstream sequences from -201 to -136 were, furthermore, capable of activating a linked β-globin gene in embryonic blood cells; however, a Gγ-globin fragment from -383 to -206 was similarly active in this assay, and the complete fragment from -383 to -136 was considerably more active than either of the smaller fragments, suggesting the presence of multiple cis-acting elements for embryonic blood cells. Our data also suggested the possibility of a negative regulatory element between -201 and -136. These results are discussed in relation to several DNA elements in the Gγ-globin upstream region, which have been shown to bind nuclear factors in erythroid cells. Finally, we observed that removal of the β-globin 3′-flanking sequences, including the 3′ enhancer, from the Gγ-globin upstream-β-globin hybrid gene resulted in a 25-fold reduction in expression in embryonic blood cells. This suggests that the β-globin 3′ enhancer is potentially active at the embryonic stage and thus cannot be solely responsible for the fetal or adult specificity of the β-globin gene.",
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