Analysis of nucleosome assembly and histone exchange using antibodies specific for acetylated H4

Perry, Carolyn A.; Dadd, Christopher A.; Allis, C. David; Annunziato, Anthony T.

doi:10.1021/bi00212a028

Cited by 54 publications

(71 citation statements)

References 65 publications

(95 reference statements)

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“…Of the histone H2A synthesized, a limited amount moves into the nucleus and is acetylated (26), and the excess unacetylated histone H2A accumulates in the cytoplasm, eventually secreted into the lumen. While the functional role of acetylation is poorly understood, it is thought to play a role in the formation of either newly synthesized or transcriptionally competent chromatin (27)(28)(29). We also observed the immunoreactivity to unacetylated histone H2A in the endoplasmic reticulum of some gastric gland cells, which would be wholly consistent with de novo histone synthesis in the cytoplasm.…”

Section: Discussionsupporting

confidence: 61%

Pepsin-Mediated Processing of the Cytoplasmic Histone H2A to Strong Antimicrobial Peptide Buforin I

Kim

Yoon

Minn

et al. 2000

The Journal of Immunology

109

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The intestinal epithelium forms a first line of innate host defense by secretion of proteins with antimicrobial activity against microbial infection. Despite the extensive studies on the antimicrobial host defense in many gastrointestinal tracts, little is known about the antimicrobial defense system of the stomach. The potent antimicrobial peptide buforin I, consisting of 39 aa, was isolated recently from the stomach tissue of an Asian toad, Bufo bufo gargarizans. In this study we examined the mechanism of buforin I production in toad stomach tissue. Buforin I is produced by the action of pepsin isozymes, named pepsin Ca and Cb, cleaving the Tyr39-Ala40 bond of histone H2A. Immunohistochemical analysis revealed that buforin I is present extracellularly on the mucosal surface, and unacetylated histone H2A, a precursor of buforin I, is localized in the cytoplasm of gastric gland cells. Furthermore, Western blot analysis showed that buforin I is also present in the gastric fluids, and immunoelectron microscopy detected localization of the unacetylated histone H2A in the cytoplasmic granules of gastric gland cells. The distinct subcellular distribution of the unacetylated histone H2A and the detection of the unacetylated buforin I both on the mucosal surface and in the lumen suggest that buforin I is produced from the cytoplasmic unacetylated histone H2A secreted into the gastric lumen and subsequently processed by pepsins. Our results indicate that buforin I along with pepsins in the vertebrate stomach may contribute to the innate host defense of the stomach against invading microorganisms.

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Section: Discussionsupporting

confidence: 61%

Pepsin-Mediated Processing of the Cytoplasmic Histone H2A to Strong Antimicrobial Peptide Buforin I

Kim

Yoon

Minn

et al. 2000

The Journal of Immunology

109

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“…To find the steady state of acetylated H3 and H4, histones of immature and mature erythrocyte chromatin fractions S150 and PE were electrophoretically resolved on AUT-polyacrylamide gels, transferred to nitrocellulose, and immunochemically stained with either antiacetylated H4 antibodies or antiacetylated H3 antibodies. Both antibodies detect the multiacetylated forms of H4 and H3, with the antibody to acetylated H3 showing a strong preference for the highest acetylated isoforms of H3 (30). We have shown previously that highly acetylated histones are found in chromatin fractions S150 and PE, but not in fraction P150 (7,24).…”

Section: State Of Acetylation Of Dynamically Acetylated Histones Inmentioning

confidence: 91%

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

Walia

Chen

Sun

et al. 1998

Journal of Biological Chemistry

104

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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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“…The antibodies against differentially modified histones were shown to specifically recognize the designated modification (Lin et al 1989;Braunstein et al 1993Braunstein et al , 1996Perry et al 1993;Lu et al 1994) and were used according to the manufacturer's recommendations (Upstate Biotechnology).…”

Section: Cytology and Microscopymentioning

confidence: 99%

A histone code in meiosis: the histone kinase, NHK-1, is required for proper chromosomal architecture in Drosophila oocytes

Ivanovska¹,

Khandan²,

Ito³

et al. 2005

Genes Dev.

114

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To promote faithful propagation of the genetic material during sexual reproduction, meiotic chromosomes undergo specialized morphological changes that ensure accurate segregation of homologous chromosomes. The molecular mechanisms that establish the meiotic chromosomal structures are largely unknown. We describe a mutation in a recently identified Histone H2A kinase, nhk-1, in Drosophila that leads to female sterility due to defects in the formation of the meiotic chromosomal structures. The metaphase I arrest and the karyosome, a critical prophase I chromosomal structure, require nucleosomal histone kinase-1 (NHK-1) function. The defects are a result of failure to disassemble the synaptonemal complex and to load condensin onto the mutant chromosomes. Embryos laid by nhk-1 −/− mutant females arrest with aberrant polar bodies and mitotic spindles, revealing that mitosis is affected as well. We analyzed the role of Histone H2A phosphorylation with respect to the histone code hypothesis and found that it is required for acetylation of Histone H3 and Histone H4 in meiosis. These studies reveal a critical role for histone modifications in chromosome dynamics in meiosis and mitosis.

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Analysis of nucleosome assembly and histone exchange using antibodies specific for acetylated H4

Cited by 54 publications

References 65 publications

Pepsin-Mediated Processing of the Cytoplasmic Histone H2A to Strong Antimicrobial Peptide Buforin I

Pepsin-Mediated Processing of the Cytoplasmic Histone H2A to Strong Antimicrobial Peptide Buforin I

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

A histone code in meiosis: the histone kinase, NHK-1, is required for proper chromosomal architecture in Drosophila oocytes

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