Transcription complex stability and chromatin dynamics in vivo

Wijgerde, Mark; Grosveld, Frank; Fraser, Peter

doi:10.1038/377209a0

Cited by 467 publications

(402 citation statements)

References 35 publications

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“…It is also possible that each cell expresses both genes from the mutant paternal allele but only one gene is expressed at a given time. The latter is analogous to the flip-flop model of gene regulation that has been proposed to explain how distal control elements allow the simultaneous expression of ␥-and ␤-globin in early development (Wijgerde et al 1995). Future studies of our mutants at the cellular level will discriminate between these possibilities.…”

Section: Discussionmentioning

confidence: 99%

Deletion of the H19 differentially methylated domain results in loss of imprinted expression of H19 and Igf2

Thorvaldsen

Duran²,

Bartolomei³

1998

Genes Dev.

615

537

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Differentially methylated sequences associated with imprinted genes are proposed to control genomic imprinting. A 2-kb region located 5 to the imprinted mouse H19 gene is hypermethylated on the inactive paternal allele throughout development. To determine whether this differentially methylated domain (DMD) is required for imprinted expression at the endogenous locus, we have generated mice harboring a 1.6-kb targeted deletion of the DMD and assayed for allelic expression of H19 and the linked, oppositely imprinted Igf2 gene. H19 is activated and Igf2 expression is reduced when the DMD deletion is paternally inherited; conversely, upon maternal transmission of the mutation, H19 expression is reduced and Igf2 is activated. Consistent with the DMD's hypothesized role of setting up the methylation imprint, the mutation also perturbs allele-specific methylation of the remaining H19 sequences. In conclusion, these experiments show that the H19 hypermethylated 5 flanking sequences are required to silence paternally derived H19.Additionally, these experiments demonstrate a novel role for the DMD on the maternal chromosome where it is required for the maximal expression of H19 and the silencing of Igf2. Thus, the H19 differentially methylated sequences are required for both H19 and Igf2 imprinting.

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Section: Discussionmentioning

confidence: 99%

Deletion of the H19 differentially methylated domain results in loss of imprinted expression of H19 and Igf2

Thorvaldsen

Duran²,

Bartolomei³

1998

Genes Dev.

615

537

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“…Probes were labeled with dinitrophenol, digoxygenin, or biotin. The oligonucleotide intron probes were as described by Wijgerde et al (1995). After hybridization, the coverslips were removed by dipping in 2× SSC, and the slides were washed three times in 2× SSC at 37°C, followed by a 5-min wash in 0.1 M Tris, …”

Section: Rnase Protectionmentioning

confidence: 99%

“…This analysis was performed as described previously (Wijgerde et al 1995). Fetal livers (13.5 day) were disrupted in PBS, and the cells were fixed onto poly-L-lysine-coated slides with 4% formaldehyde/5% acetic acid in normal saline for 20 min at room temperature.…”

Section: Rnase Protectionmentioning

confidence: 99%

“…At this stage in development, ∼20% of loci transcribe the human ␥ genes and the remaining 80% transcribe the ␤ gene (Wijgerde et al 1996). Double-label primary transcript in situ hybridization was performed with gene-specific intron probes (Wijgerde et al 1995) and a probe specific to either the human LCR or G ␥-globin 3Ј-flanking region (equivalent to NRO probes G10-G14). In addition, the ␦-␤-intergenic region and ␤ 3Ј-flanking region (equivalent to NRO probes B11 and B12; see below) were investigated for transcriptional activity (Fig.…”

Section: Detection Of Lcr and Intergenic Transcription In Erythroid Tmentioning

confidence: 99%

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Intergenic transcription and transinduction of the human β-globin locus

Ashe¹,

Monks²,

Wijgerde³

et al. 1997

Genes Dev.

Self Cite

320

238

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We have identified novel nuclear transcripts in the human ␤-globin locus using nuclear run-on analysis in erythroid cell lines and in situ hybridization analysis of erythroid tissue. These transcripts extend across the LCR and intergenic regions but are undetectable in nonerythroid cells. Surprisingly, transient transfection of a ␤-globin gene (⑀, ␥, or ␤) induces transcription of the LCR and intergenic regions from the chromosomal ␤-globin locus in nonerythroid cell lines. The ␤-globin genes themselves, however, remain transcriptionally silent. Induction is dependent on transcription of the globin gene in the transfected plasmid but does not require protein expression. Using in situ hybridization analysis, we show that the plasmid colocalizes with the endogenous ␤-globin locus providing insight into the mechanism of transinduction.

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“…The idea of a dynamic equilibrium between the active and the inactive state of a gene locus, which (in the case of transgenes) is a function of the genomic integration site, is supported by a variety of studies. In a series of elegant experiments, Fraser and co-workers (27) have demonstrated that the human ␤-globin LCR-promoter interaction is dynamic and switches between several promoters of the downstream located globin genes (25). The authors also analyzed certain transgenic mouse lines carrying deletion mutants of the complete ␤-globin locus, which render the transgene susceptible to genomic position effects.…”

Section: Incomplete Gene Loci Do Not Form Stable Transcription Complexesmentioning

confidence: 99%

The Chicken Lysozyme Locus as a Paradigm for the Complex Developmental Regulation of Eukaryotic Gene Loci

Bonifer

Jägle²,

Huber

1997

Journal of Biological Chemistry

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Gene loci of higher organisms have complex structural features. In some cases their coding regions occupy many, even hundreds, of kilobases of DNA. Additionally, the sequences that contain the information for the correct spatial and temporal regulation of a particular gene locus during development often exceed the extensions of the coding region by severalfold.The question of what type of information is encoded in these vast amounts of DNA has puzzled researchers from the beginning.It is now clear that eukaryotic genes are regulated by a number of different cis-regulatory elements distributed over large distances. A convenient way to assay the number and the distribution of cis-regulatory elements has been the mapping of DHS 1 in chromatin. Such local chromatin perturbations are in most cases caused by the binding of transcription factors to their cognate DNA sequences. The pattern of DHS can undergo dramatic developmental changes, indicating a change in the activity of cis-regulatory elements. In addition, the analysis of protein-DNA interactions at a single-nucleotide resolution level in vivo has demonstrated that, depending on the developmental stage, different combinations of transcription factors can occupy the same cis-regulatory element (1, 2). These experiments indicate that the transcriptional activation of a gene locus is achieved by the cooperation of several different cisregulatory elements, which, in turn, assemble transcription factors in a sequential, developmentally controlled fashion. However, the assembly of active transcription factor complexes on natural genes does not occur on naked DNA but in a chromatin context, where nucleosome-DNA interactions have to be counteracted. Hence, the activation of a gene locus requires at least the following steps: the perturbation of chromatin structure by the binding of transcription factors on cis-regulatory elements, the developmentally controlled reorganization of transcription factor complexes, the assembly of the basal transcription machinery and its interaction with upstream regulatory elements, the onset of mRNA synthesis, and, in many cases, the maintenance of an active transcriptional state during multiple rounds of DNA synthesis.How can the molecular basis of locus activation be experimentally studied? While the basal activities of individual cisregulatory elements of particular gene loci can be analyzed by transient and stable transfection experiments, the molecular mechanism of activation of a gene locus from the transcriptionally silent state can only be studied in a developing system, preferentially in transgenic animals. The ideal model locus should be small, thus facilitating the manipulation of individual cis-regulatory elements within the context of an entire genomic locus, and it should be extensively characterized on the molecular level. In addition, to dissect the role of different cis-regulatory elements in the developmental control of gene locus activation, it should be possible to follow cell differentiation experimentally, thus enab...

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Transcription complex stability and chromatin dynamics in vivo

Cited by 467 publications

References 35 publications

Deletion of the H19 differentially methylated domain results in loss of imprinted expression of H19 and Igf2

Deletion of the H19 differentially methylated domain results in loss of imprinted expression of H19 and Igf2

Intergenic transcription and transinduction of the human β-globin locus

The Chicken Lysozyme Locus as a Paradigm for the Complex Developmental Regulation of Eukaryotic Gene Loci

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