Mapping of linear histone regions exposed at the surface of the nucleosome in solution

Stemmer, Christian; Briand, Jean‐Paul; Muller, Sylviane

doi:10.1006/jmbi.1997.1270

Cited by 43 publications

(42 citation statements)

References 32 publications

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“…The buforin I-specific Ab was immunologically unreactive with respect to both nuclear and cytoplasmic histone H2As. A mapping study of the linear histone H2A regions revealed that the peptide sequence of the synthetic congener buforin I is not localized on the surface of histone H2A protein, and therefore histone H2A is not accessible to the buforin I Ab (25). A mAb (BWA3) to unacetylated histone H2A was used for the detection of histone H2A in the cytoplasm.…”

Section: Discussionmentioning

confidence: 99%

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

Kim

Yoon

Minn

et al. 2000

The Journal of Immunology

111

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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: Discussionmentioning

confidence: 99%

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

Kim

Yoon

Minn

et al. 2000

The Journal of Immunology

111

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“…Primary antibodies were 1) mouse monoclonal anti-Oct-4 (2 g/ml; Santa Cruz Biotechnology, Santa Cruz, CA), 2) goat anti-secreted protein which is acidic and rich in cysteines (SPARC) (2 g/ml; R&D Systems, Minneapolis, MN), 3) rabbit anti-histone H3 (COOH-terminal 130 -135 peptide; 1/1000 -1/3000; Stemmer et al, 1997), and 4) mouse monoclonal antipolymerase (7C2; 1/10,000 dilution; Besse et al, 1995). Secondary antibodies were 1) donkey anti-mouse IgG (heavy and light chains) conjugated with horseradish peroxidase (HRP; 0.16 g/ml; Jackson ImmunoResearch Laboratories, West Grove, PA; used with anti-polymerase), 2) sheep anti-mouse IgG conjugated with HRP (1/4000; GE Healthcare; used with anti-Oct-4), 3) donkey anti-goat IgG (heavy and light chains) conjugated with HRP (0.16 g/ml; Jackson ImmunoResearch Laboratories), and 4) donkey anti-rabbit IgG (heavy and light chains) conjugated with HRP (0.16 g/ml; Jackson ImmunoResearch Laboratories).…”

Section: Immunoblottingmentioning

confidence: 99%

A Conserved Organization of Transcription during Embryonic Stem Cell Differentiation and in Cells with High C Value

Faro-Trindade

Cook

2006

MBoC

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Although we have detailed information on the alterations occurring in steady-state levels of all cellular mRNAs during differentiation, we still know little about more global changes. Therefore, we investigated the numbers of molecules of RNA polymerase II that are active-and the way those molecules are organized-as two mouse cells (aneuploid F9 teratocarcinoma, and euploid and totipotent embryonic stem cells) differentiate into parietal endoderm. Quantitative immunoblotting shows the number of active molecules roughly halves. Transcription sites (detected by light and electron microscopy after allowing engaged polymerases to extend nascent transcripts in bromouridine-triphosphate) are uniformly distributed throughout the nucleoplasm. The numbers of such sites fall during differentiation as nuclei become smaller, but site density and diameter remain roughly constant. Similar site densities and diameters are found in salamander (amphibian) cells with 11-fold larger genomes, and in aneuploid HeLa cells. We conclude that active polymerases and their nascent transcripts are concentrated in a limited number of discrete nucleoplasmic sites or factories, and we speculate that the organization of transcription is conserved during both differentiation and evolution to a high C value. INTRODUCTIONDifferentiation involves the repression of some genes and the activation of others. For example, during erythropoiesis nuclei become smaller and heterochromatic as many genes are silenced; concurrently, globin genes become active by associating with specific nuclear sites known as "factories" or "active chromatin hubs" (Cook, 1999(Cook, , 2002de Laat and Grosveld, 2003;Osborne et al., 2004). Although microarrays provide detailed information on changes in steady-state mRNA levels during differentiation (Kelly and Rizzino, 2000;Harris and Childs, 2002;Tanaka et al., 2002;Sperger et al., 2003), there is little information on how primary rates of transcription change. For example, we do not know by how much global rates vary as any mammalian cell differentiates. Eukaryotic genomes also differ in size more than 200,000-fold in a way unrelated to organismal complexity; there is no consensus as to the basis of this C-value enigma (Hartl, 2000;Gregory, 2001), or on how transcription might be organized in such differently sized genomes.Against this background, we investigated the changes that occur in the rates and organization of transcription as two mouse cells differentiate. Mouse F9 teratocarcinoma cells are a well-studied aneuploid line that can be induced to differentiate into parietal endoderm, which is characterized by flattened cells that secrete tissue-type plasminogen activator, laminin, and type IV collagen (Strickland and Mahdavi, 1978;Strickland et al., 1980;Hogan et al., 1983). ESF/48 -1 cells are totipotent embryonic stem (ES) cells with a normal karyotype; when transferred into host blastocysts, they contribute to all tissues (including germ lines) of the resulting chimeras (Brook and Gardner, 1997), and they can be induc...

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“…Nucleosomes were prepared, as described previously (30), from mouse liver, and purified on a 5-29% (w/v) sucrose gradient. The nucleosome preparations were characterized by 1.5% agarose gel electrophoresis, and the content of histones was checked by 18% SDS-PAGE (29).…”

Section: Peptides Histones and Nucleosomesmentioning

confidence: 99%

“…The conditions used for the coating of native DNA and individual histones were described previously (30). In each assay, sera were also tested in a noncoated well incubated with coating buffer alone as a control.…”

Section: Elisa For Ab Measurementsmentioning

confidence: 99%

“…They encompassed residues 10-33 of H2B, 34-48 of H2A, 85-99 and 94-108 of H3, as well as 16-39 (14 -28), [49][50][51][52][53][54][55][56][57][58][59][60][61][62][63], and 71-94 of H4. Interestingly, most of these T cell epitopes are located in regions of histones that have been shown previously to be readily accessible at the surface of nucleosome (30). In the present study, we tested 11 peptides from H4 and 15 peptides from H3 for recognition by CD4 ϩ T cells from unprimed (NZB ϫ New Zealand White (NZW))F 1 mice (BW; H-2 d/z ), a classical lupus-prone mouse model that was not previously investigated for reactivity of anti-histone T cells.…”

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confidence: 92%

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CD4+ T Cells from (New Zealand Black × New Zealand White)F1 Lupus Mice and Normal Mice Immunized Against Apoptotic Nucleosomes Recognize Similar Th Cell Epitopes in the C Terminus of Histone H3

Fournel

Neichel

Dali

et al. 2003

The Journal of Immunology

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We have previously reported that peptide 88-99 of histone H4 represents a minimal T cell epitope recognized by Th cells from nonautoimmune BALB/c (H-2d/d) mice immunized with nucleosomes. In this study, we tested a panel of overlapping peptides spanning the whole sequences of H4 and H3 for recognition by CD4+ T cells from unprimed (New Zealand Black (NZB) × New Zealand White (NZW))F1 lupus mice (H-2d/z). None of the 11 H4 peptides was recognized by CD4+ T cells from (NZB × NZW)F1 mice. In contrast, these cells proliferated and secreted IL-2, IL-10, and IFN-γ upon ex vivo stimulation with H3 peptides representing sequences 53-70, 64-78, and 68-85. Peptides 56-73 and 61-78 induced the production of IFN-γ and IL-10, respectively, without detectable proliferation, suggesting that they may act as partial agonist of the TCR. Th cells from unprimed BALB/c mice and other lupus-prone mice such as SNF1 (H-2d/q) and MRL/lpr (H-2k/k) mice did not recognize any peptides present within the H3 region 53-85. We further demonstrated that immunization of normal BALB/c mice with syngeneic liver nucleosomes and spleen apoptotic cells, but not with nonapoptotic syngeneic cells, induced Th cell responses against several peptides of the H3 region 53-85. Moreover, we found that this conserved region of H3, which is accessible at the surface of nucleosomes, is targeted by Abs from (NZB × NZW)F1 mice and lupus patients, and contains motifs recognized by several distinct HLA-DR molecules. It might thus be important in the self-tolerance breakdown in lupus.

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Mapping of linear histone regions exposed at the surface of the nucleosome in solution

Cited by 43 publications

References 32 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

A Conserved Organization of Transcription during Embryonic Stem Cell Differentiation and in Cells with High C Value

CD4+ T Cells from (New Zealand Black × New Zealand White)F1 Lupus Mice and Normal Mice Immunized Against Apoptotic Nucleosomes Recognize Similar Th Cell Epitopes in the C Terminus of Histone H3

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