Activation of β-major globin gene transcription is associated with recruitment of NF-E2 to the β-globin LCR and gene promoter

Sawado, Tomoyuki; Igarashi, Kazuhiko; Groudine, Mark

doi:10.1073/pnas.181344198

Cited by 121 publications

(173 citation statements)

References 48 publications

(48 reference statements)

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“…Immunoprecipitated DNA was analyzed by PCR using primers specific for the human p53 gene promoter. For the semi-quantitative ChIP assay, duplex-PCR was performed as described (Sawado et al, 2001). Primer sequences used for ChIP assays are available by request.…”

Section: Chip Assaymentioning

confidence: 99%

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Epigenetic regulation of the human p53 gene promoter by the CTCF transcription factor in transformed cell lines

Soto-Reyes

Recillas‐Targa

2010

Oncogene

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Section: Chip Assaymentioning

confidence: 99%

“…Primer sequences used for ChIP assays are available by request. For the duplex PCR we used for normalization b-actin for the close histone marks and MyoD for open histone marks (Sawado et al, 2001). For each antibody, at least three independent ChIP experiments were performed.…”

Section: Chip Assaymentioning

confidence: 99%

Epigenetic regulation of the human p53 gene promoter by the CTCF transcription factor in transformed cell lines

Soto-Reyes

Recillas‐Targa

2010

Oncogene

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“…p45 NF-E2, which contains a transactivation domain, interacts with TAFII130 (Amrolia et al 1997) and CBP in vitro (Cheng et al 1997;Hung et al 2001) and is required for pol II recruitment to the ␤ maj globin gene promoter in mouse erythroleukemia (MEL) cells (Johnson et al 2001). Recently, we reported that NF-E2 binds to both the LCR and the ␤-globin promoter region, even though no canonical NF-E2 binding sites are present in the promoter (Sawado et al 2001). This observation has reinforced speculation that the LCR functions through interactions with the promoter.…”

mentioning

confidence: 99%

“…To gain insight into which transcription events are influenced by the LCR, we have performed ChIP analyses of histone modifications and factor recruitment at the adult ␤-globin gene in two model systems: (1) differentiation of 745A MEL cells, a process accompanied by a greater than 100-fold increase in ␤-globin gene expression (Sawado et al 2001); and (2) primary erythroid cells derived from ⌬LCR/WT heterozygous mice.…”

mentioning

confidence: 99%

The β-globinlocus control region (LCR) functions primarily by enhancing the transition from transcription initiation to elongation

Sawado¹,

Halow²,

Bender³

et al. 2003

Genes Dev.

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164

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To investigate the molecular basis of ␤-globin gene activation, we analyzed factor recruitment and histone modification at the adult ␤-globin gene in wild-type (WT)/locus control region knockout (⌬LCR) heterozygous mice and in murine erythroleukemia (MEL) cells. Although histone acetylation and methylation (Lys 4) are high before and after MEL differentiation, recruitment of the erythroid-specific activator NF-E2 to the promoter and preinitiation complex (PIC) assembly occur only after differentiation. We reported previously that targeted deletion of the LCR reduces ␤-globin gene expression to 1%-4% of WT without affecting promoter histone acetylation. Here, we report that NF-E2 is recruited equally efficiently to the adult ␤-globin promoters of the ⌬LCR and WT alleles. Moreover, the LCR deletion reduces PIC assembly only twofold, but has a dramatic effect on Ser 5 phosphorylation of RNA polymerase II and transcriptional elongation. Our results suggest at least three distinct stages in ␤-globin gene activation: (1) [Keywords: Allele-specific chromatin immunoprecipitation analysis; locus control region; ␤-globin locus; transcription elongation; NF-E2; CTD-phosphorylation] Supplemental material is available at http://www.genesdev.org. Received January 6, 2003; revised version accepted February 19, 2003. In higher eukaryotes, gene activation involves several events including chromatin opening, activator binding to regulatory regions, recruitment of basal transcription factors (TFIIA to TFIIJ) and RNA polymerase II (pol II) to the promoter [preinitiation complex (PIC) assembly], and transcription elongation. A general model has emerged in which activators function to stabilize or modulate transcription through interactions with histone acetyltransferases, as well as components of the basal transcription machinery, such as TATA binding protein (TBP), TBP-associated factors (TAFs), and TFIIB (for review, see Lemon and Tjian 2000). PIC assembly is followed by initiation and elongation, during which the Cterminal domain (CTD) of the RPB1 subunit of pol II is phosphorylated (for review, see Dahmus 1996). These steps in gene activation are often regulated by distal elements called enhancers (for review, see Blackwood and Kadonaga 1998;Martin 2001). Enhancers increase either the rate of transcription, the number of the templates engaged in transcription, or both. Studies using transgenes or in vitro templates have linked enhancer activities to various events such as PIC assembly (Kim et al. 1998;Yie et al. 1999), histone acetylation at the promoter (Madisen et al. 1998), and nuclear localization (Francastel et al. 1999). However, it is poorly understood which events in transactivation are the actual targets of long-range enhancer function at native gene loci.The murine ␤-globin gene locus is a model system for studying the molecular mechanisms of enhancer-dependent gene activation at a native locus. The locus contains embryonic and adult ␤-like globin genes that are ordered as they are expressed during development. Hig...

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“…However, in uninduced MEL cells, it is maintained at the basal level but the locus is still kept open without detectable change from the state of induced cells. (33,34) After opening the domain, the distal enhancer elements further augment transcription. Hence, the strandbreaking model does not exclude the looping or linking model.…”

Section: Chromatin Remodeling and Chromatin Domain Openingmentioning

confidence: 99%

A hypothesis for chromatin domain opening

Liu

Ling

2003

BioEssays

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SummaryThe eukaryotic genome is organized into different domains by cis-acting elements, such as boundaries/ insulators and matrix attachment regions, and is packaged with different degrees of condensation. In the M phase, the chromatin becomes further highly condensed into chromosomes. The first step for transcriptional activation of a given gene, at a particular time during development, in any locus, is the opening of its chromatin domain. This locus needs to be kept in this state in each early G 1 phase during every cell cycle. Certain distal enhance elements, including locus control regions (LCRs) and enhancers, are believed to perform this target chromatin domain opening process and several models have been proposed to explain distal enhance action. But they did not explain precisely how a given chromatin domain is opened. Based on various studies, we propose a hypothesis for the mechanism of opening chromatin on a large scale. One important mechanism may involved breaking one or two DNA strands and reducing the linking numbers within chromatin domain. The topological changes can overpass some complexes formed on DNA strands and can be transmitted from specific localized points over a broad region, until boundary elements or insulators are reached. These may initiate downstream events such as propagation of histone acetylation and the binding of transcription factors to proximal promoters and may further augment the action mediated by distal enhancer elements.

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Activation of β-major globin gene transcription is associated with recruitment of NF-E2 to the β-globin LCR and gene promoter

Cited by 121 publications

References 48 publications

Epigenetic regulation of the human p53 gene promoter by the CTCF transcription factor in transformed cell lines

Epigenetic regulation of the human p53 gene promoter by the CTCF transcription factor in transformed cell lines

The β-globinlocus control region (LCR) functions primarily by enhancing the transition from transcription initiation to elongation

A hypothesis for chromatin domain opening

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