Main regulatory element (MRE) of the Danio rerio α/β-globin gene domain exerts enhancer activity toward the promoters of the embryonic-larval and adult globin genes

2016 ◽  
Vol 50 (6) ◽  
pp. 900-908 ◽  
Author(s):  
A. P. Kovina ◽  
N. V. Petrova ◽  
S. V. Razin ◽  
O. V. Yarovaia
Keyword(s):  
Danio Rerio ◽  
Regulatory Element ◽  
Globin Gene ◽  
Globin Genes ◽  
Enhancer Activity ◽  
Globin Gene Domain

The coordinate replication of the human beta-globin gene domain reflects its transcriptional activity and nuclease hypersensitivity

1988 ◽  
Vol 8 (11) ◽  
pp. 4958-4965
Author(s):  
V Dhar ◽  
D Mager ◽  
A Iqbal ◽  
C L Schildkraut
Keyword(s):  
Cell Lines ◽  
Dna Sequences ◽  
Globin Gene ◽  
K562 Cells ◽  
Globin Genes ◽  
Beta Globin ◽  
Beta Globin Gene ◽  
Hypersensitive Sites ◽  
Flanking Sequences ◽  
Globin Gene Domain

The temporal order of replication of DNA sequences in the chromosomal domain containing the human beta-globin gene cluster and its flanking sequences (140 kilobases) was measured and compared in two different human cell lines. In human erythroleukemia (K562) cells, in which embryonic and fetal globin genes are transcribed, all of the sequences we examined from the beta-globin domain replicated early during S phase, while in HeLa cells, in which globin genes are transcriptionally silent, these sequences replicated late during S. Potential sites of initiation of DNA replication within this domain were identified. The beta-globin gene domain was also found to differ with respect to the nuclease sensitivity of the chromatin in these two cell lines. In K562 cells, hypersensitive sites for endogenous nucleases and DNase I were present in the chromatin near the earliest-replicating segments in the beta-globin domain.

scholarly journals Characterization of the DNase I hypersensitive site 3' of the human beta globin gene domain

Blood ◽  
1993 ◽  
Vol 81 (10) ◽  
pp. 2781-2790
Author(s):  
DE Fleenor ◽  
RE Kaufman
Keyword(s):  
Topoisomerase Ii ◽  
Globin Gene ◽  
Dnase I ◽  
Globin Genes ◽  
Beta Globin ◽  
Hypersensitive Site ◽  
Mobility Shift ◽  
Enhancer Activity ◽  
Beta Globin Gene

The members of the human beta globin gene family are flanked by strong DNase I hypersensitive sites. The collection of sites 5' to the epsilon globin gene is able to confer high levels of expression of linked globin genes, but a function has not been assigned to the site 3' to the beta globin gene (3'HS1). Our analysis of this DNase I super hypersensitive site shows that the region is composed of multiple DNase I sites. By examination of the DNA sequence, we have determined that the region is very A/T-rich and contains topoisomerase II recognition sequences, as well as several consensus binding motifs for GATA-1 and AP-1/NF-E2. Gel mobility shift assays indicate that the region can interact in vitro with GATA-1 and AP-1/NF-E2, and functional studies show that the region serves as a scaffold attachment region in both erythroid and nonerythroid cell lines. Whereas many of the physical features of 3'HS1 are shared by 5'HS2 (a component of the 5' locus control region), transient expression studies show that 3' HS1 does not share the erythroid-specific enhancer activity exhibited by 5'HS2.

Deletion of the mouse α-globin regulatory element (HS −26) has an unexpectedly mild phenotype

Blood ◽  
2002 ◽  
Vol 100 (10) ◽  
pp. 3450-3456 ◽  
Author(s):  
Eduardo Anguita ◽  
Jacqueline A. Sharpe ◽  
Jacqueline A. Sloane-Stanley ◽  
Cristina Tufarelli ◽  
Douglas R. Higgs ◽  
...  
Keyword(s):  
Regulatory Element ◽  
Globin Gene ◽  
Mean Corpuscular Hemoglobin ◽  
Single Element ◽  
Surprising Result ◽  
Globin Genes ◽  
Mild Phenotype ◽  
Mild Disease ◽  
Expression Studies ◽  
High Level

Natural deletions of the region upstream of the human α-globin gene cluster, together with expression studies in cell lines and transgenic mice, identified a single element (HS −40) as necessary and perhaps sufficient for high-level expression of the α-globin genes. A similar element occupies the corresponding position upstream of the mouse (m) α-globin genes (mHS −26) and was thought to have similar functional properties. We knocked out mHS −26 by homologous recombination and observed the surprising result that instead of the expected severe α-thalassemia phenotype, the mice had a mild disease. Transcription levels of the mouse genes were reduced by about 50%, but homozygotes were healthy, with normal hemoglobin levels and only mild decreases in mean corpuscular volume and mean corpuscular hemoglobin. These results may indicate differences in the regulation of the α-globin clusters in mice and humans or that additionalcis-acting elements remain to be characterized in one or both clusters.

Mapping of structural and transcription-related matrix attachment sites in the alpha-globin gene domain of avian erythroblasts and erythrocytes

1990 ◽  
Vol 10 (10) ◽  
pp. 5349-5358
Author(s):  
G Farache ◽  
S V Razin ◽  
J Rzeszowska-Wolny ◽  
J Moreau ◽  
F R Targa ◽  
...  
Keyword(s):  
Nuclear Matrix ◽  
Large Fraction ◽  
Globin Gene ◽  
Dna Interaction ◽  
Globin Genes ◽  
Origin Of Replication ◽  
Attachment Sites ◽  
Alpha Globin ◽  
Globin Gene Domain ◽  
Structural Matrix

The positions of preferential DNA interaction with the nuclear matrix were mapped within the domain of the chicken alpha-globin genes in transcriptionally active erythroblast nuclei and inactive nuclei of mature erythrocytes. In the latter, only two major distinct attachment sites were observed, close to the A + T-rich sequences previously found at the boundaries of the domain. Sequencing of these structural matrix attachment points revealed several known DNA motifs; some of them were present on both sides of the domain. In actively transcribing erythroblast nuclei of adult animals, a large fraction of the transcribed area was represented in nuclear matrix DNA, including upstream and downstream elements. In particular, adult alpha A- and alpha D-globin genes were found in matrix DNA, while the transcribed but translationally unexpressed embryonic pi gene was underrepresented. The data are discussed in terms of the existence of stable or structural and expression-related matrix attachment sites; correlations to the origin of replication and the units of transcription of the domain are shown.

Developmental regulation of the human embryonic beta-like globin gene is mediated by synergistic interactions among multiple tissue- and stage-specific elements

1993 ◽  
Vol 13 (12) ◽  
pp. 7457-7468
Author(s):  
W L Trepicchio ◽  
M A Dyer ◽  
M H Baron
Keyword(s):  
Developmental Regulation ◽  
Regulatory Element ◽  
Globin Gene ◽  
Negative Control ◽  
Erythroid Cell ◽  
Globin Genes ◽  
Erythroid Cells ◽  
Specific Expression ◽  
Systematic Analysis ◽  
Specific Regulation

The stage-specific regulation of mammalian embryonic globin genes has been an experimentally elusive problem, in part because of the developmentally early timing of their expression. We have carried out a systematic analysis of truncation and internal deletion mutations within the 5'-flanking region of the human embryonic beta-like globin gene (epsilon) in erythroid and nonerythroid cell lines. Within a 670-bp region upstream from the constitutive promoter are multiple positive and negative control elements. Of these, a positive regulatory element (epsilon-PRE II) which is active only in embryonic erythroid cells is of particular interest. Remarkably, although it is inactive on its own, in the presence of other sequences located further upstream, it confers tissue- and developmental stage-specific expression on a constitutive epsilon-globin or heterologous promoter. The activity of epsilon-PRE II is also modulated by another positive regulatory domain located further downstream to direct erythroid cell-specific, but little or no embryonic stage-specific, transcription. A nuclear factor highly enriched in embryonic erythroid cells binds specifically within a 19-bp region of epsilon-PRE II. Nuclei from adult erythroid cells also contain a factor that binds to this region but forms a complex of faster electrophoretic mobility. We speculate that interactions between epsilon-PRE II and other upstream control elements play an important role in the developmental regulation of the human embryonic beta-like globin gene.

scholarly journals The coordinate replication of the human beta-globin gene domain reflects its transcriptional activity and nuclease hypersensitivity.

1988 ◽  
Vol 8 (11) ◽  
pp. 4958-4965 ◽  
Author(s):  
V Dhar ◽  
D Mager ◽  
A Iqbal ◽  
C L Schildkraut
Keyword(s):  
Cell Lines ◽  
Dna Sequences ◽  
Globin Gene ◽  
K562 Cells ◽  
Globin Genes ◽  
Beta Globin ◽  
Beta Globin Gene ◽  
Hypersensitive Sites ◽  
Flanking Sequences ◽  
Globin Gene Domain

The temporal order of replication of DNA sequences in the chromosomal domain containing the human beta-globin gene cluster and its flanking sequences (140 kilobases) was measured and compared in two different human cell lines. In human erythroleukemia (K562) cells, in which embryonic and fetal globin genes are transcribed, all of the sequences we examined from the beta-globin domain replicated early during S phase, while in HeLa cells, in which globin genes are transcriptionally silent, these sequences replicated late during S. Potential sites of initiation of DNA replication within this domain were identified. The beta-globin gene domain was also found to differ with respect to the nuclease sensitivity of the chromatin in these two cell lines. In K562 cells, hypersensitive sites for endogenous nucleases and DNase I were present in the chromatin near the earliest-replicating segments in the beta-globin domain.

scholarly journals The major regulatory element upstream of the alpha-globin gene has classical and inducible enhancer activity

Blood ◽  
1993 ◽  
Vol 81 (4) ◽  
pp. 1058-1066 ◽  
Author(s):  
S Ren ◽  
XN Luo ◽  
GF Atweh
Keyword(s):  
Messenger Rna ◽  
Phorbol Esters ◽  
Regulatory Element ◽  
Globin Gene ◽  
Expression System ◽  
K562 Cells ◽  
Erythroid Differentiation ◽  
Enhancer Activity ◽  
Alpha Globin ◽  
Functional Features

Abstract A major positive regulatory element has recently been identified 40 kb upstream from the human zeta 2-globin gene. This regulatory element increases the expression of a linked alpha-globin gene in mouse erythroleukemia cells and in transgenic mice. This element has been shown to share many of the structural and functional features of the locus control region (LCR) of the beta-globin gene cluster. We have examined the activity of a small fragment from this regulatory domain (alpha LCR) in a transient expression system. We show that this element is active as an enhancer in the erythroid environment of K562 cells. It is somewhat less effective as an enhancer in the nonerythroid environment of HeLa cells. This alpha LCR fragment does not exhibit promoter specificity because it can activate both the promoter of its endogenous target gene and the heterologous promoter of the SV40 early genes. Although the major activity of this element is mediated by its interaction with the promoter of the alpha-globin gene, some increase in activity is seen when structural elements from the 5′ end of the alpha-globin gene are included with the target promoter. In addition, we show that the enhancing activity of the alpha LCR is potentiated by hemin-induction of K562 cells. Whereas phorbol esters that induce megakaryocytic differentiation of K562 cells markedly decrease alpha- globin messenger RNA accumulation, they do not seem to have a negative effect on the activity of the alpha LCR. These studies suggest a role for the alpha LCR in the basal activity of the alpha-globin gene in erythroid cells and in its increased expression seen with erythroid differentiation. The mechanism of negative regulation of alpha-globin gene expression in phorbol-differentiated K562 cells does not appear to be mediated through the action of the alpha LCR.

scholarly journals The mouse alpha-globin locus regulatory element

Blood ◽  
1995 ◽  
Vol 86 (2) ◽  
pp. 766-775 ◽  
Author(s):  
G Gourdon ◽  
JA Sharpe ◽  
DR Higgs ◽  
WG Wood
Keyword(s):  
Regulatory Element ◽  
Globin Gene ◽  
Globin Genes ◽  
Band Shift ◽  
Core Element ◽  
Protein Binding Sites ◽  
Endogenous Gene ◽  
Human Counterpart ◽  
Alpha Globin ◽  
High Level

We have identified and cloned the major alpha globin locus regulatory element in the mouse (m alpha RE). This element shows a high level of sequence homology to its human counterpart (HS -40) and lies between the same two exons of an upstream, widely expressed gene in both species. Footprinting and band shift studies of the core element show conservation of many (but not all) of the protein binding sites identified as functionally important in HS -40. The functional equivalence of the mouse element was shown by attaching it to a human alpha globin gene and examining expression in transgenic mice. Readily detectable levels of human alpha mRNA were produced in these mice but they were lower than the endogenous gene expression and did not show copy number dependence. These results suggest that sequences additional to this major regulatory element may be necessary to obtain complete regulation of the alpha globin genes in both species.

scholarly journals A DNA-bending protein interacts with an essential upstream regulatory element of the human embryonic beta-like globin gene.

1996 ◽  
Vol 16 (3) ◽  
pp. 829-838 ◽  
Author(s):  
M A Dyer ◽  
R Naidoo ◽  
R J Hayes ◽  
C J Larson ◽  
G L Verdine ◽  
...  
Keyword(s):  
Regulatory Element ◽  
Globin Gene ◽  
Gene Promoter ◽  
Developmental State ◽  
Biochemical Properties ◽  
Erythroid Cell ◽  
Control Element ◽  
Specific Gene ◽  
Globin Genes ◽  
Specific Expression

The mammalian beta-like globin gene family has served as an important model system for analysis of tissue- and developmental state-specific gene regulation. Although the activities of a number of regulatory proteins have been implicated in the erythroid cell-specific transcription of globin genes, the mechanisms that restrict their expression to discrete stages of development are less well understood. We have previously identified a novel regulatory element (PRE II) upstream from the human embryonic beta-like globin gene (epsilon) that synergizes with other sequences to confer tissue- and stage-specific expression on a minimal epsilon-globin gene promoter in cultured embryonic erythroid cells. Binding of an erythroid nuclear protein (PRE II-binding factor [PRE-IIBF]) to the PRE II control element is required for promoter activation. Here we report on some of the biochemical properties of PREIIBF, including the characterization of its specificity and affinity for DNA. The embryonic and adult forms of PREIIBF recognize their cognate sequences with identical specificities, supporting our earlier conclusion that they are very similar proteins. PREIIBF binds DNA as a single polypeptide with an Mr of approximately 80,000 to 85,000 and introduces a bend into the target DNA molecule. These results suggest a mechanism by which PREIIBF may contribute to the regulation of the embryonic beta-like globin gene within the context of a complex locus.

scholarly journals Mapping of structural and transcription-related matrix attachment sites in the alpha-globin gene domain of avian erythroblasts and erythrocytes.

1990 ◽  
Vol 10 (10) ◽  
pp. 5349-5358 ◽  
Author(s):  
G Farache ◽  
S V Razin ◽  
J Rzeszowska-Wolny ◽  
J Moreau ◽  
F R Targa ◽  
...  
Keyword(s):  
Nuclear Matrix ◽  
Large Fraction ◽  
Globin Gene ◽  
Dna Interaction ◽  
Globin Genes ◽  
Origin Of Replication ◽  
Attachment Sites ◽  
Alpha Globin ◽  
Globin Gene Domain ◽  
Structural Matrix

The positions of preferential DNA interaction with the nuclear matrix were mapped within the domain of the chicken alpha-globin genes in transcriptionally active erythroblast nuclei and inactive nuclei of mature erythrocytes. In the latter, only two major distinct attachment sites were observed, close to the A + T-rich sequences previously found at the boundaries of the domain. Sequencing of these structural matrix attachment points revealed several known DNA motifs; some of them were present on both sides of the domain. In actively transcribing erythroblast nuclei of adult animals, a large fraction of the transcribed area was represented in nuclear matrix DNA, including upstream and downstream elements. In particular, adult alpha A- and alpha D-globin genes were found in matrix DNA, while the transcribed but translationally unexpressed embryonic pi gene was underrepresented. The data are discussed in terms of the existence of stable or structural and expression-related matrix attachment sites; correlations to the origin of replication and the units of transcription of the domain are shown.

Sign in / Sign up

Export Citation Format

Share Document