Conserved elements containing NF-E2 and tandem GATA binding sites are required for erythroid-specific chromatin structure reorganization within the human beta-globin locus control region

Pomerantz, Oded; Goodwin, Andrew; Joyce, T.; Lowrey, Christopher H.

doi:10.1093/nar/26.24.5684

Cited by 26 publications

(18 citation statements)

References 33 publications

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“…In contrast to the ε-globin promoter, which is structurally altered by the action of the HS2 enhancer, the enhancer could potentially form its distinctive nuclease-sensitive structure in chromatin in an autonomous fashion (48). Alternatively, an enhancer may not be sufficient to form a disrupted chromatin structure in the absence of a promoter (52).…”

Section: Resultsmentioning

confidence: 99%

Structural and Functional Cross-Talk between a Distant Enhancer and the ɛ-Globin Gene Promoter Shows Interdependence of the Two Elements in Chromatin

McDowell

Dean

1999

Molecular and Cellular Biology

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We investigated the requirements for enhancer-promoter communication by using the human ␤-globin locus control region (LCR) DNase I-hypersensitive site 2 (HS2) enhancer and the -globin gene in chromatinized minichromosomes in erythroid cells. Activation of globin genes during development is accompanied by localized alterations of chromatin structure, and CACCC binding factors and GATA-1, which interact with both globin promoters and the LCR, are believed to be critical for globin gene transcription activation. We found that an HS2 element mutated in its GATA motif failed to remodel the -globin promoter or activate transcription yet HS2 nuclease accessibility did not change. Accessibility and transcription were reduced at promoters with mutated GATA-1 or CACCC sites. Strikingly, these mutations also resulted in reduced accessibility at HS2. In the absence of a globin gene, HS2 is similarly resistant to nuclease digestion. In contrast to observations in Saccharomyces cerevisiae, HS2-dependent promoter remodeling was diminished when we mutated the TATA box, crippling transcription. This mutation also reduced HS2 accessibility. The results indicate that the -globin promoter and HS2 interact both structurally and functionally and that both upstream activators and the basal transcription apparatus contribute to the interaction. Further, at least in this instance, transcription activation and promoter remodeling by a distant enhancer are not separable.A central question in developmental biology is how enhancers activate gene transcription in a tissue-and developmentalstage-specific fashion, a complex process which takes place in the chromatin environment of the nucleus. We have addressed this issue by studying components of the human ␤-globin gene locus, the locus control region (LCR) DNase I-hypersensitive site 2 (HS2) enhancer and the embryonic ε-globin gene. The locus consists of five genes expressed sequentially during development and a multicomponent, far-upstream regulatory element, the LCR (for a review, see reference 6). In naturally occurring thalassemias with large deletions encompassing the LCR and upstream sequences, expression of the downstream globin genes is abolished, and the chromatin structure of the locus becomes resistant to DNase I cleavage (20). Therefore, it has long been thought that the LCR mediates both decondensation of the chromatin of the globin locus and activation of transcription of the genes at different stages in development. However, recent experiments in which the mouse or human ␤-globin LCR was deleted in its natural chromosomal context demonstrate that while the LCR is required for high-level transcription, it may not be required for decondensation of the locus or correct developmental regulation of the globin genes (16, 51). Thus, the LCR, at a minimum, fulfills the role of a traditional enhancer.The LCR contains four HS sites (HS1 to HS4) detected exclusively in the chromatin of erythroid cells (21, 55). Of these, only HS2 has enhancer activity in transient transfection assays, a...

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Section: Resultsmentioning

confidence: 99%

Structural and Functional Cross-Talk between a Distant Enhancer and the ɛ-Globin Gene Promoter Shows Interdependence of the Two Elements in Chromatin

McDowell

Dean

1999

Molecular and Cellular Biology

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“…Our own studies on HS4, HS3, and HS2, in which we have stably transfected mutant HS binding sites into MEL cells, indicate that loss of conserved NF-E2-and tandem, inverted GATA-binding elements significantly reduces or eliminates completely the DNase sensitivity of the HSs, particularly when both elements are mutated within the same construct (14,15). Boyes and Felsenfeld (27) performed similar experiments using chicken ␤-globin 3Ј enhancer sequences.…”

Section: Roles Of Nf-e2 Gata-1 Eklf and Sp1 In Lcr Hsmentioning

confidence: 85%

“…In contrast to the promoterassociated HSs of the locus, the ␤-globin LCR HSs are ϳ10 -20-fold more sensitive to DNase I digestion (9,13,14). At the nucleosomal level, the LCR cores are characterized by regions of nucleosome disruption or displacement spanning ϳ200 bp (14,15). At the DNA level, each LCR HS core sequence is highly conserved between species (16).…”

mentioning

confidence: 99%

“…We have previously observed that the cores of LCR HSs 1-4 contain tandem, inverted GATA-binding elements located ϳ50 bp downstream from AP-1/NF-E2-binding elements (also known as maf response elements) (14,17). These binding sites are located within the nucleosome-free regions of the HS cores; when these sites are mutated, loss of the normal chromatin structure of the HS and of appropriate expression from linked transgenes in transgenic mice occurs (8,14,18). Recent experiments in ␤-globin yeast artificial chromosome (YAC) transgenic mice have emphasized the importance of the HS core elements to the function of the LCR, because deletion of a 280-bp element containing the HS4 core or a 225-bp element containing the HS3 core both result in dramatic changes in gene expression despite the presence of otherwise intact ␤-globin yeast artificial chromosomes (YACs) (19,20).…”

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confidence: 99%

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In Vivo Formation of a Human β-Globin Locus Control Region Core Element Requires Binding Sites for Multiple Factors Including GATA-1, NF-E2, Erythroid Kruppel-like Factor, and Sp1

Goodwin

McInerney

Glander

et al. 2001

Journal of Biological Chemistry

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The active elements of the ␤-globin locus control region (LCR) are located within domains of unique chromatin structure. These nuclease hypersensitive sites (HSs) are characterized by high DNase I sensitivity, erythroid specificity, similar nucleosomal structure, and evolutionarily conserved clusters of cis-acting elements that are required for the formation and function of the core elements. To determine the requirements for HS core formation in the setting of nuclear chromatin, we constructed a series of artificial HS cores containing binding sites for GATA-1, NF-E2, and Sp1. In contrast to the results of previous in vitro experiments, we found that when constructs were stably integrated in mouse erythroleukemia cells the binding sites for NF-E2, GATA-1, or Sp1 alone or in any combination were unable to form core HS structures. We subsequently identified two new cis-acting elements from the LCR HS4 core that, when combined with the NF-E2, Sp1, and tandem inverted GATA elements, result in core structure formation. Both new cis-acting elements bind Sp1, and one binds erythroid Kruppel-like factor (EKLF). We conclude that in vivo ␤-globin LCR HS core formation is more complex than previously thought and that several factors are required for this process to occur.

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“…11A). The second site that we examined for histone acetylation was DNase I-hypersensitive site 3 (HS3) of the LCR, since it contains GATA-1-binding sites that are essential for HS formation (59). While histone H3 acetylation increased approximately twofold in control MEL cells, no change in histone H3 acetylation was observed in both MEL-PU.1 clones (Fig.…”

Section: Pu1 Inhibits Differentiation-induced Histone Acetylation Atmentioning

confidence: 99%

Inhibition of CBP-Mediated Protein Acetylation by the Ets Family Oncoprotein PU.1

Wang

Kim

et al. 2002

Molecular and Cellular Biology

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Aberrant expression of PU.1 inhibits erythroid cell differentiation and contributes to the formation of murine erythroleukemias (MEL). The molecular mechanism by which this occurs is poorly understood. Here we show that PU.1 specifically and efficiently inhibits CBP-mediated acetylation of several nuclear proteins, including the hematopoietic transcription factors GATA-1, NF-E2, and erythroid Krüppel-like factor. In addition, PU.1 blocks acetylation of histones and interferes with acetylation-dependent transcriptional events. CBP acetyltransferase activity increases during MEL cell differentiation as PU.1 levels decline and is inhibited by sustained PU.1 expression. Finally, PU.1 inhibits the differentiation-associated increase in histone acetylation at an erythroid-specific gene locus in vivo. Together, these findings suggest that aberrant expression of PU.1 and possibly other members of the Ets family of oncoproteins subverts normal cellular differentiation in part by inhibiting the acetylation of critical nuclear factors involved in balancing cellular proliferation and maturation.Cellular transformation can result from deregulated expression of nuclear transcription factors. The Ets family oncoprotein PU

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Conserved elements containing NF-E2 and tandem GATA binding sites are required for erythroid-specific chromatin structure reorganization within the human beta-globin locus control region

Cited by 26 publications

References 33 publications

Structural and Functional Cross-Talk between a Distant Enhancer and the ɛ-Globin Gene Promoter Shows Interdependence of the Two Elements in Chromatin

Structural and Functional Cross-Talk between a Distant Enhancer and the ɛ-Globin Gene Promoter Shows Interdependence of the Two Elements in Chromatin

In Vivo Formation of a Human β-Globin Locus Control Region Core Element Requires Binding Sites for Multiple Factors Including GATA-1, NF-E2, Erythroid Kruppel-like Factor, and Sp1

Inhibition of CBP-Mediated Protein Acetylation by the Ets Family Oncoprotein PU.1

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