scholarly journals Effect of deletion of 5'HS3 or 5'HS2 of the human beta-globin locus control region on the developmental regulation of globin gene expression in beta-globin locus yeast artificial chromosome transgenic mice.

1996 ◽  
Vol 93 (13) ◽  
pp. 6605-6609 ◽  
Author(s):  
K. R. Peterson ◽  
C. H. Clegg ◽  
P. A. Navas ◽  
E. J. Norton ◽  
T. G. Kimbrough ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transgenic Mice ◽  
Control Region ◽  
Yeast Artificial Chromosome ◽  
Developmental Regulation ◽  
Globin Gene ◽  
Artificial Chromosome ◽  
Locus Control Region ◽  
Beta Globin ◽  
Globin Gene Expression

scholarly journals A single beta-globin locus control region element (5' hypersensitive site 2) is sufficient for developmental regulation of human globin genes in transgenic mice.

1992 ◽  
Vol 12 (5) ◽  
pp. 2057-2066 ◽  
Author(s):  
B J Morley ◽  
C A Abbott ◽  
J A Sharpe ◽  
J Lida ◽  
P S Chan-Thomas ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transgenic Mice ◽  
Control Region ◽  
Developmental Regulation ◽  
Globin Gene ◽  
Locus Control Region ◽  
Erythroid Cell ◽  
Globin Genes ◽  
Beta Globin ◽  
Hypersensitive Site

The beta-globin gene complex is regulated by an upstream locus control region (LCR) which is responsible for high-level, position-independent, erythroid-cell-specific expression of the genes in the cluster. Its role in the developmental regulation of beta-like globin gene transcription remains to be established. We have examined the effect of a single LCR element, hypersensitive site 2 (HS2), on the developmental regulation of the human fetal gamma and adult beta genes in transgenic mice. In mice bearing HS2A gamma beta and HS2G gamma A gamma-117 delta beta human globin gene constructs, switching from gamma- to beta-gene expression begins at about day 13.5 of gestation and is largely completed shortly after birth. The larger construct also demonstrates a switch in G gamma- to A gamma-gene expression during the gamma-to-beta switch similar to that observed during normal human development. We conclude that HS2 alone is sufficient for developmental regulation of the human beta-globin genes.

A single beta-globin locus control region element (5' hypersensitive site 2) is sufficient for developmental regulation of human globin genes in transgenic mice

1992 ◽  
Vol 12 (5) ◽  
pp. 2057-2066
Author(s):  
B J Morley ◽  
C A Abbott ◽  
J A Sharpe ◽  
J Lida ◽  
P S Chan-Thomas ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transgenic Mice ◽  
Control Region ◽  
Developmental Regulation ◽  
Globin Gene ◽  
Locus Control Region ◽  
Erythroid Cell ◽  
Globin Genes ◽  
Beta Globin ◽  
Hypersensitive Site

The beta-globin gene complex is regulated by an upstream locus control region (LCR) which is responsible for high-level, position-independent, erythroid-cell-specific expression of the genes in the cluster. Its role in the developmental regulation of beta-like globin gene transcription remains to be established. We have examined the effect of a single LCR element, hypersensitive site 2 (HS2), on the developmental regulation of the human fetal gamma and adult beta genes in transgenic mice. In mice bearing HS2A gamma beta and HS2G gamma A gamma-117 delta beta human globin gene constructs, switching from gamma- to beta-gene expression begins at about day 13.5 of gestation and is largely completed shortly after birth. The larger construct also demonstrates a switch in G gamma- to A gamma-gene expression during the gamma-to-beta switch similar to that observed during normal human development. We conclude that HS2 alone is sufficient for developmental regulation of the human beta-globin genes.

scholarly journals Silencing of Aγ-Globin Gene Expression during Adult Definitive Erythropoiesis Mediated by GATA-1-FOG-1-Mi2 Complex Binding at the −566 GATA Site

2008 ◽  
Vol 28 (10) ◽  
pp. 3101-3113 ◽  
Author(s):  
Susanna Harju-Baker ◽  
Flávia C. Costa ◽  
Halyna Fedosyuk ◽  
Renee Neades ◽  
Kenneth R. Peterson
Keyword(s):  
Gene Expression ◽  
Transgenic Mice ◽  
Yeast Artificial Chromosome ◽  
Fetal Liver ◽  
Globin Gene ◽  
Protein A ◽  
Artificial Chromosome ◽  
Nurd Complex ◽  
Definitive Erythropoiesis ◽  
Globin Gene Expression

ABSTRACTAutonomous silencing of γ-globin transcription is an important developmental regulatory mechanism controlling globin gene switching. An adult stage-specific silencer of theAγ-globin gene was identified between −730 and −378 relative to the mRNA start site. A marked copy of theAγ-globin gene inserted between locus control region 5′ DNase I-hypersensitive site 1 and the ε-globin gene was transcriptionally silenced in adult β-globin locus yeast artificial chromosome (β-YAC) transgenic mice, but deletion of the 352-bp region restored expression. This fragment reduced reporter gene expression in K562 cells, and GATA-1 was shown to bind within this sequence at the −566 GATA site. Further, the Mi2 protein, a component of the NuRD complex, was observed in erythroid cells with low γ-globin levels, whereas only a weak signal was detected when γ-globin was expressed. Chromatin immunoprecipitation of fetal liver tissue from β-YAC transgenic mice demonstrated that GATA-1, FOG-1, and Mi2 were recruited to theAγ-globin −566 orGγ-globin −567 GATA site when γ-globin expression was low (day 18) but not when γ-globin was expressed (day 12). These data suggest that during definitive erythropoiesis, γ-globin gene expression is silenced, in part, by binding a protein complex containing GATA-1, FOG-1, and Mi2 at the −566/−567 GATA sites of the proximal γ-globin promoters.

Long-Range Enhancement of β-Globin Gene Expression Is Dependent on Gt Motifs Residing in the HS3 Core Element of the Locus Control Region.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1222-1222
Author(s):  
Patrick A. Navas ◽  
Andrew B. Stergachis ◽  
Hadar H. Sheffer ◽  
Xin Ye ◽  
Mary Stafford ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transgenic Mice ◽  
Control Region ◽  
Regulatory Element ◽  
Globin Gene ◽  
Locus Control Region ◽  
Core Element ◽  
Rnase Protection ◽  
Definitive Erythropoiesis ◽  
Globin Gene Expression

Abstract Normal expression of the human β-globin genes is dependent on a powerful regulatory element residing upstream of the globin gene cluster referred as the locus control region (LCR), physically characterized by five DNAseI-hypersensitive sites (HS1 to HS5). Of particular interest is HS3, which is characterized by a 225 bp core element region containing seven GT motifs (GT1 to GT7) that alternate with four GATA-1 binding sites. The GT motifs are bound by a family of Kruppel-like factors and are important for proper expression of many housekeeping and tissue-specific genes. We previously demonstrated in transgenic mice carrying a 248 kb human β-globin yeast artificial chromosome in which the GT6 motif (GT6m β-YAC) was mutated resulted in a decrease in ε- and γ-globin gene expression during embryonic erythropoiesis and a decrease in γ-globin expression during definitive erythropoiesis, thus, providing evidence that a single transcriptional motif distantly located can have profound effects on gene expression. We have used the same β-YAC transgenic mouse model system to analyze the function of the remaining six GT motifs of the HS3 core element. Currently we have produced transgenic mice carrying either a mutation of GT1/2 (GT1 and GT2 motifs overlap) or GT3. Total RNA was isolated from yolk sac, liver and blood samples of transgenic F2 embryos at 12- and 14-day postconception and adult blood, and subjected to RNAse protection analysis. The GT1/2m β-YAC transgenic mice exhibited normal ε- and γ-globin gene expression during embryonic erythropoiesis, but during definitive erythropoiesis γ-globin gene expression was reduced 4 to 5 fold in day 12- and day 14- fetal livers. In the two GT1/2m β-YAC transgenic lines produced, β-globin gene expression was reduced and levels varied from 12.3% ± 2.9% to 63.2% ± 5.8% of endogenouse murine α-globin indicating that β-globin expression is strongly influenced by the position of integration of the transgene. In contrast, the GT3m β-YAC transgenic mice the mutation to GT3 had no effect on globin gene expression during development. Our results suggest that multiple GT motifs within the same highly complex regulatory element contribute differently to the enhancement of the downstream genes.

Reduced beta-Globin Gene Expression in Adult Mice Containing Deletions of Locus Control Region 5' HS-2 or 5' HS-3a

1998 ◽  
Vol 850 (1 COOLEY'S ANEM) ◽  
pp. 45-53 ◽  
Author(s):  
TIMOTHY J. LEY ◽  
BRUCE HUG ◽  
STEVEN FIERING ◽  
ELLIOT EPNER ◽  
M. A. BENDER ◽  
...  
Keyword(s):  
Gene Expression ◽  
Control Region ◽  
Globin Gene ◽  
Locus Control Region ◽  
Beta Globin ◽  
Beta Globin Gene ◽  
Adult Mice ◽  
Globin Gene Expression

scholarly journals Linear Distance from the Locus Control Region Determines ε-Globin Transcriptional Activity

2007 ◽  
Vol 27 (16) ◽  
pp. 5664-5672 ◽  
Author(s):  
Motoshi Shimotsuma ◽  
Hitomi Matsuzaki ◽  
Osamu Tanabe ◽  
Andrew D. Campbell ◽  
James Douglas Engel ◽  
...  
Keyword(s):  
Gene Expression ◽  
Control Region ◽  
Yeast Artificial Chromosome ◽  
Globin Gene ◽  
Artificial Chromosome ◽  
Locus Control Region ◽  
Globin Genes ◽  
Control Of Gene Expression ◽  
Linear Distance ◽  
Globin Promoter

ABSTRACT Enhancer elements modulate promoter activity over vast chromosomal distances, and mechanisms that ensure restrictive interactions between promoters and enhancers are critical for proper control of gene expression. The human β-globin locus control region (LCR) activates expression of five genes in erythroid cells, including the proximal embryonic ε- and the distal adult β-globin genes. To test for possible distance sensitivity of the genes to the LCR, we extended the distance between the LCR and genes by 2.3 kbp within the context of a yeast artificial chromosome, followed by the generation of transgenic mice (TgM). In these TgM lines, ε-globin gene expression decreased by 90%, while the more distantly located γ- or β-globin genes were not affected. Remarkably, introduction of a consensus EKLF binding site into the ε-globin promoter rendered its expression distance insensitive; when tested in an EKLF-null genetic background, expression of the mutant ε-globin gene was severely compromised. Thus, the ε-globin gene differs in its distance sensitivity to the LCR from the other β-like globin genes, which is, at least in part, determined by the transcription factor EKLF.

Deletions within the Mouse β-Globin Locus Control Region Preferentially Reduce βmin Globin Gene Expression

Genomics ◽  
2000 ◽  
Vol 63 (3) ◽  
pp. 417-424 ◽  
Author(s):  
Raouf Alami ◽  
M.A. Bender ◽  
Yong-Qing Feng ◽  
Steven N. Fiering ◽  
Bruce A. Hug ◽  
...  
Keyword(s):  
Gene Expression ◽  
Control Region ◽  
Globin Gene ◽  
Locus Control Region ◽  
Globin Gene Expression

Direct interaction of NF-E2 with hypersensitive site 2 of the β-globin locus control region in living cells

Blood ◽  
2000 ◽  
Vol 96 (1) ◽  
pp. 334-339 ◽  
Author(s):  
E. Camilla Forsberg ◽  
Karen M. Downs ◽  
Emery H. Bresnick
Keyword(s):  
Gene Expression ◽  
Control Region ◽  
Globin Gene ◽  
Locus Control Region ◽  
Erythroid Cell ◽  
Hypersensitive Site ◽  
Intact Cells ◽  
Erythroleukemia Cells ◽  
Murine Erythroleukemia ◽  
Globin Gene Expression

The human β-globin locus control region (LCR) confers high-level, tissue-specific expression to the β-globin genes. Tandem Maf recognition elements (MAREs) within the hypersensitive site 2 (HS2) subregion of the LCR are important for the strong enhancer activity of the LCR. Multiple proteins are capable of interacting with these sites in vitro, including the erythroid cell- and megakaryocyte-specific transcription factor, NF-E2. The importance of NF-E2 for β-globin gene expression is evident in murine erythroleukemia cells lacking the p45 subunit of NF-E2. These CB3 cells have a severe defect in - and β-globin gene transcription, which can be restored by expression of NF-E2. However, mice nullizygous for p45 express nearly normal levels of β-globin. Thus, either a redundant factor(s) exists in mice that can functionally replace NF-E2, or NF-E2 does not function through the LCR to regulate β-globin gene expression. To address this issue, we asked whether NF-E2 binds directly to the tandem MAREs of HS2 in intact cells. Using a chromatin immunoprecipitation assay, we provide evidence for NF-E2 binding directly and specifically to HS2 in living erythroleukemia cells and in mouse fetal liver. The specific immunoisolation of HS2 sequences was dependent on the presence of p45 and on intact MAREs within HS2. These results support a direct role for NF-E2 in the regulation of β-globin gene expression through activation of the LCR.

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