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

McDowell, Jennifer C.; Dean, Ann

doi:10.1128/mcb.19.11.7600

Cited by 23 publications

(21 citation statements)

References 65 publications

(89 reference statements)

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“…There is also evidence for communications between the ε gene promoter and HS2. Mutation of GATA-1 or CACCC site in the ε promoter resulted in reduced accessibility at HS2, and HS2-dependent promoter remodeling was diminished when the ε gene TATA box was mutated (22). Our results add to this evidence by showing that a ␥ CCAAT mutation can affect the in vivo recruitment of GATA-1 and CBP/ p300 to HS2 and HS3.…”

Section: Discussionsupporting

confidence: 72%

Role of NF-Y in In Vivo Regulation of the γ-Globin Gene

Duan

Stamatoyannopoulos

2001

Molecular and Cellular Biology

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The duplicated CCAAT box is required for ␥ gene expression. We report here that the transcriptional factor NF-Y is recruited to the duplicated CCAAT box in vivo. A mutation of the duplicated CCAAT box that severely disrupts the NF-Y binding also reduces the accessibility level of the ␥ gene promoter, affects the assembly of basal transcriptional machinery, and increases the recruitment of GATA-1 to the locus control region (LCR) and the proximal promoter and the recruitment of transcription cofactor CBP/p300 to the LCR. These findings suggest that recruitment of NF-Y to the duplicated CCAAT box plays a role in the chromatin opening of the ␥ gene promoter as well as in the communication between the ␥ gene promoter and the LCR.

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Section: Discussionsupporting

confidence: 72%

Role of NF-Y in In Vivo Regulation of the γ-Globin Gene

Duan

Stamatoyannopoulos

2001

Molecular and Cellular Biology

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“…These sites (see Fig. 1 A) include the conserved GATA-1 site at Ϫ165, the CACC motif at Ϫ110, the TATA box (39), and a ϩ6 GATA-1 motif within a putative initiator element of the core promoter (49). The mutant genes were linked to a wild-type HS2 located 2.2 kb upstream of the promoter.…”

Section: Resultsmentioning

confidence: 99%

“…Clustered point mutations were introduced by site-directed mutagenesis into HS2 or the -globin promoter to eliminate binding of NF-E2, GATA-1, and CACC factors. The mutations, transfection conditions, and culture conditions of K562 cells have been described (29,39). Clones studied in this work stably carried 10-40 copies of minichromosomes.…”

Section: Methodsmentioning

confidence: 99%

“…The minichromosome model allows high-resolution structural analyses of the chromatin transitions that accompany transcription activation in organisms as diverse as yeast and humans (37,38). Transcription activation of a 4-Kb embryonic -globin gene on minichromosomes in K562 cells is HS2-dependent, and is accompanied by DNase I HS formation, increased restriction enzyme accessibil-ity, and histone hyperacetylation at the promoter (39,40). Here we show that enhancer-dependent remodeling results in nucleosome mobilization at the -globin promoter, and, like histone hyperacetylation, requires the HS2 MAREs.…”

mentioning

confidence: 99%

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A major role for the TATA box in recruitment of chromatin modifying complexes to a globin gene promoter

Gui

Dean²

2003

Proc. Natl. Acad. Sci. U.S.A.

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The developmentally regulated mammalian ␤-globin genes are activated by a distant locus control region͞enhancer. To understand the role of chromatin remodeling complexes in this activation, we used stably replicated chromatin templates, in which transcription activation of the human embryonic -globin gene depends on the tandem Maf-recognition elements (MAREs) within the ␤-globin locus control region HS2 enhancer, to which the erythroid factor NF-E2 binds. The HS2 MAREs are required for nucleosome mobilization and histone hyperacetylation at the distant promoter. Nucleosome mobilization also requires the promoter TATA box, and is independent of histone hyperacetylation. In contrast, promoter hyperacetylation requires the promoter GATA-1, and CACC-factor activator motifs, as well as the TATA box. ChIP analysis reveals that NF-E2 is associated with the active -globin promoter, which lacks an NF-E2 binding sequence, in a TATA box and HS2͞MARE-dependent fashion. NF-E2 association with the -globin promoter coincides with that of RNA polymerase II at both regulatory sites. The results emphasize MARE-TATA box interactions in the recruitment of complexes modifying promoter chromatin for transcription activation and imply close physical interaction between widely separated regulatory sequences mediated through these sites.

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“…Clustered point mutations were introduced into HS2 to eliminate binding of NF-E2. These mutations, the transfection conditions, the growth of K562 cells, and individual clones carrying minichromosomes have been described (20,29). A representative clone of each type was studied in this work.…”

Section: Methodsmentioning

confidence: 99%

Acetylation of a Specific Promoter Nucleosome Accompanies Activation of the ɛ-Globin Gene by β-Globin Locus Control Region HS2

Gui

Dean

2001

Molecular and Cellular Biology

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On stably replicating episomes, transcriptional activation of the -globin promoter by the ␤-globin locus control region HS2 enhancer is correlated with an increase in nuclease sensitivity which is limited to the TATAproximal nucleosome (N1). To elucidate what underlies this increase in nuclease sensitivity and the link between chromatin modification and gene expression, we examined the nucleoprotein composition and histone acetylation status of transcriptionally active and inactive promoters. Micrococcal nuclease digestion of active promoters in nuclei released few nucleosome-like nucleoprotein complexes containing N1 sequences in comparison to results with inactive promoters. We also observed that N1 DNA fragments from active promoters are of a subnucleosomal length. Nevertheless, chromatin immunoprecipitation experiments indicate that histones H3 and H4 are present on N1 sequences from active promoters, with H3 being dramatically hyperacetylated compared with that from inactive promoters and vector sequences. Strikingly, H3 in the adjacent upstream nucleosome (N2) does not appear to be differentially acetylated in active and inactive promoters, indicating that the nucleosome modification of the promoter that accompanies transactivation by HS2 is highly directed and specific. However, global acetylation of histones in vivo by trichostatin A did not activate transcription in the absence of HS2, suggesting that HS2 contributes additional activities necessary for transactivation. N1 sequences from active promoters also contain reduced levels of linker histone H1. The detection of a protected subnucleosomal sized N1 DNA fragment and the recovery of N1 DNA sequences in immunoprecipitations using antiacetylated H3 and H4 antibodies argue that N1 is present, but in an altered conformation, in the active promoters.

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

Cited by 23 publications

References 65 publications

Role of NF-Y in In Vivo Regulation of the γ-Globin Gene

Role of NF-Y in In Vivo Regulation of the γ-Globin Gene

A major role for the TATA box in recruitment of chromatin modifying complexes to a globin gene promoter

Acetylation of a Specific Promoter Nucleosome Accompanies Activation of the ɛ-Globin Gene by β-Globin Locus Control Region HS2

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