Chicken erythrocyte polynucleosomes which are soluble at physiological ionic strength and contain linker histones are highly enriched hi β-globin gene sequences

Ridsdale, Jenna; Davie, James R.

doi:10.1093/nar/15.3.1081

Cited by 99 publications

(79 citation statements)

References 27 publications

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“…Nucleosome Preparation and Histone Isolation-Chicken immature erythrocyte salt-soluble chromatin S150 was prepared as described previously (20). The salt-soluble chromatin fragments are enriched in acetylated core histones and will remain soluble in the kinase buffer, which contains 10 mM MgCl 2 .…”

Section: Methodsmentioning

confidence: 99%

Phosphorylation of Histones by Tissue Transglutaminase

Mishra

Saleh

Espino

et al. 2006

Journal of Biological Chemistry

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Tissue transglutaminase 2 (TG2) has recently been shown to have intrinsic serine/threonine kinase activity. Since histones are known to be cross-linked by TG2, we investigated whether histones are also substrates for TG2 kinase activity. TG2 was able to phosphorylate H1, H2A, H2B, H3, and H4 histones in vitro. Using peptide substrates and phosphospecific antibodies we demonstrated that TG2 phosphorylated Ser 10 in H3 and that this phosphorylation was reduced by acetylation, whereas phosphorylation of Ser 10 by TG2 enhanced acetylation. Furthermore we demonstrated that exogenous TG2 phosphorylated H1 and H3 in nucleosome preparations. We examined the abundance of TG2 in DNA-associated proteins from MCF-7 cells treated with phorbol ester (TPA) and 17␤-estradiol (E2). TG2 abundance was significantly reduced in E2-treated cells and enhanced in TPA-treated cells. In summary we have demonstrated that TG2 is able to phosphorylate purified histone proteins, and H3 and H1 in chromatin preparations, and it is associated with chromatin in breast cancer cells. These studies suggest a novel role for TG2 in the regulation of chromatin structure and function. Tissue transglutaminase 2 (TG2)4 is a ubiquitously expressed, calcium-dependent trans-amidating acyltransferase that cross-links proteins resulting in polymerization (1). In addition, it has other less characterized functions including as a protein-disulfide isomerase (2), as a G protein-coupled membrane receptor (3) and as a serine/threonine kinase (4). It is localized to many compartments in the cell including the cell membrane, the cytoplasm, and the nucleus (1). TG2 is involved in a multitude of cellular process of which the best studied is apoptosis where it is responsible for the formation of apoptotic bodies.TG2 is translocated to the nucleus by a mechanism that involves importin-␣3 (5, 6), where it can cross-link histones (7,8). Cross-linking of the core histone subunits, H2A and H2B, by TG2 appears to involve glutamine and lysine residues at COOH-terminal domain (9). Histone H1, has also been shown to be a substrate for TG2 (9). Histones bind in a sequence-independent manner to DNA to form chromatin. They are subject to extensive post-translational modification that appears to be important in regulating chromatin function. For example, the aminoterminal tails of histones can be both phosphorylated and acetylated and these modifications are thought to regulate chromatin structure to facilitate transcription, DNA replication, mitosis, and DNA repair.Histone cross-linking by TG2 may be responsible for the changes in DNA function such as transcription and replication (11), although the exact molecular mechanisms responsible have not been elucidated, and many apparently contradictory data exist. For example, it appears that TG2 can only cross-link histones when released from the nucleosome. Since cross-linking of histone by TG2 does not occur when the histones were organized in nucleosome (9), it is not immediately obvious how this cross-linking activity of TG2 could ...

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Section: Methodsmentioning

confidence: 99%

Phosphorylation of Histones by Tissue Transglutaminase

Mishra

Saleh

Espino

et al. 2006

Journal of Biological Chemistry

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“…The mono-, di-, tri-, and tetra-acetylated isoforms of H4 are marked as 1, 2, 3, and 4, respectively. Note that the content of H1 and H5 in fraction S150 is typically lower than that of the other chromatin fractions (65). (4); the net result is a low steady state level of highly acetylated histone isoforms in untreated immature cells.…”

Section: State Of Acetylation Of Dynamically Acetylated Histones Inmentioning

confidence: 97%

Histone Acetylation Is Required to Maintain the Unfolded Nucleosome Structure Associated with Transcribing DNA

Walia

Chen

Sun

et al. 1998

Journal of Biological Chemistry

106

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Nucleosomes associated with transcribing chromatin of mammalian cells have an unfolded structure in which the normally buried cysteinyl-thiol group of histone H3 is exposed. In this study we analyzed transcriptionally active/competent DNA-enriched chromatin fractions from chicken mature and immature erythrocytes for the presence of thiol-reactive nucleosomes using organomercury-agarose column chromatography and hydroxylapatite dissociation chromatography of chromatin fractions labeled with [ 3 H]iodoacetate. In mature and immature erythrocytes, the active DNA-enriched chromatin fractions are associated with histones that are rapidly highly acetylated and rapidly deacetylated. When histone deacetylation was prevented by incubating cells with histone deacetylase inhibitors, sodium butyrate or trichostatin A, thiol-reactive H3 of unfolded nucleosomes was detected in the soluble chromatin and nuclear skeleton-associated chromatin of immature, but not mature, erythrocytes. We did not find thiol-reactive nucleosomes in active DNA-enriched chromatin fractions of untreated immature erythrocytes that had low levels of highly acetylated histones H3 and H4 or in chromatin of immature cells incubated with inhibitors of transcription elongation. This study shows that transcription elongation is required to form, and histone acetylation is needed to maintain, the unfolded structure of transcribing nucleosomes.Acetylation of the core histones (H2A, H2B, H3, and H4) is a dynamic process catalyzed by histone acetyltransferases and histone deacetylases (1, 2). In chicken immature erythrocytes, 4% of the modifiable lysine sites participate in dynamic histone acetylation. These core histones are rapidly acetylated (t1 ⁄2 ϭ 12 min for monoacetylated H4) and rapidly deacetylated (t1 ⁄2 ϭ 5 min for the tetraacetylated isoform of H4) (3, 4). Histones undergoing rapid acetylation and deacetylation are associated with transcriptionally active chromatin (5-7). The recent findings that histone acetyltransferases and deacetylases are transcriptional coactivators and corepressors have increased our understanding of how the process of dynamic histone acetylation is established on transcriptionally active chromatin (2, 8).Transcriptionally active chromatin has a soluble and insoluble nature (9). Transcribed DNA is found in chromatin fragments that are soluble in 0.15 M NaCl and/or 2 mM MgCl 2 and in chromatin fragments associated with the low salt-insoluble residual nuclear material (nuclear skeletons) (for review, see Davie (10)). Chromatin engaged in transcription is thought to be retained by the nuclear skeleton by multiple dynamic attachments between the nuclear matrix and transcribed chromatin; hence rendering the transcribing chromatin insoluble (11, 12). As histone acetyltransferase and deacetylase activities are associated with the nuclear matrix (7, 13), we proposed that these nuclear matrix-bound enzymes may mediate some of the dynamic attachments between active chromatin and nuclear matrix (13,14). Most information on the structure ...

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“…It has been suggested that acetylation of H4 during nucleosome assembly regulates the binding of H1 and the ability of chromatin to condense (34,35). While in some cases active genes are hyperacetylated and contain H1 (10,31,37), it has also been reported that while H1 binds to acetylated oligonucleosomes, this binding inhibits transcription (53). In addition, studies have demonstrated that histone acetylation alters the capacity of histone H1 to condense chromatin (36) and that the presence of H1 affects the ability of transcription factors to interact with the DNA (19,39).…”

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confidence: 99%

Histone H1 Is a Specific Repressor of Core Histone Acetylation in Chromatin

Herrera¹,

West²,

Schiltz

et al. 2000

Molecular and Cellular Biology

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Although a link between histone acetylation and transcription has been established, it is not clear how acetylases function in the nucleus of the cell and how they access their targets in a chromatin fiber containing H1 and folded into a highly condensed structure. Here we show that the histone acetyltransferase (HAT) p300/CBP-associated factor (PCAF), either alone or in a nuclear complex, can readily acetylate oligonucleosomal substrates. The linker histones, H1 and H5, specifically inhibit the acetylation of mono-and oligonucleosomes and not that of free histones or histone-DNA mixtures. We demonstrate that the inhibition is due mainly to steric hindrance of H3 by the tails of linker histones and not to condensation of the chromatin fiber. Cellular PCAF, which is complexed with accessory proteins in a multiprotein complex, can overcome the linker histone repression. We suggest that linker histones hinder access of PCAF, and perhaps other HATs, to their target acetylation sites and that perturbation of the linker histone organization in chromatin is a prerequisite for efficient acetylation of the histone tails in nucleosomes.Chromatin, with its associated linker histones, is a highly condensed structure that constrains the genome into the nucleus of the cell and suppresses various DNA-related activities such as transcription and replication. Transcriptional activation has been associated with changes in the structure of both chromatin and nucleosomes (57, 58). These changes are mediated by chromatin remodeling complexes (59) and by reversible modification of histones (46,56). Indeed, there is a strong correlation between the acetylation state of core histones and the transcriptional competence of specific genes (21, 46, 52). This correlation has been strengthened by the finding that several transcription factors have intrinsic histone acetyltransferase (HAT) activity (28,46) and that mutants lacking HAT activity fail to activate transcription of their target genes (23,55). Recent studies suggest that HATs function in the context of multiprotein complexes in vivo and that the acetylase activity of these complexes is more efficient than that of the isolated transcription factors (15,32,48). It is conceivable that some of the proteins found in these multiprotein complexes function to facilitate histone acetylation in the context of chromatin.In chromatin, the N-terminal tails of the core histones are thought to be involved in internucleosomal interactions and have been shown to be required for formation of higher-order, condensed chromatin structure (3,12,17). Studies using oligonucleosomes condensed with salt indicate that the HAT GCN5 can efficiently acetylate the N-terminal tail of histone H3 (51), suggesting that at least some of the acetylation targets are available in condensed chromatin. An additional major factor, known to be involved in the formation and stabilization of a higher-order, condensed chromatin structure, is histone H1. Numerous studies have demonstrated that the presence of H1 inhibits transc...

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Chicken erythrocyte polynucleosomes which are soluble at physiological ionic strength and contain linker histones are highly enriched hi β-globin gene sequences

Cited by 99 publications

References 27 publications

Phosphorylation of Histones by Tissue Transglutaminase

Phosphorylation of Histones by Tissue Transglutaminase

Histone Acetylation Is Required to Maintain the Unfolded Nucleosome Structure Associated with Transcribing DNA

Histone H1 Is a Specific Repressor of Core Histone Acetylation in Chromatin

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