Use of histone antibodies for studying chromatin topography and the phosphorylation of chromatin subunits.

Muller, Sylviane; Mazen, A.; Martinage, Arlette; Regenmortel, M.H.V. Van

doi:10.1002/j.1460-2075.1984.tb02150.x

Cited by 47 publications

(12 citation statements)

References 62 publications

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“…In agreement with our earlier data (Huletsky et al, 1989;Muller et al, 1982Muller et al, , 1984Thibeault et al, 1992), only a few histone antibody probes reacted with nucleosomes in this ELISA format; namely, the four antisera induced against the complete core histones, the MAb LG2-2 reacting with the N terminus 1-25 of H2B and only a few antisera directed against histone peptides; namely, antisera to peptides 65-85 of H2A (rabbit Dub), 118-135 (rabbit Mar) and 130-135 of H3 (rabbits Giz and Mel; Table 2). We found no signi®cant difference in terms of absorbance values measured in ELISA when mono-, di-and trinucleosomes or long chains (n 20 to 35) of chromatin were used for coating plates.…”

Section: Reactivity Of Histone Antibodies With Nucleosomes In a Direcsupporting

confidence: 94%

“…In native chromatin, the regions corresponding to residues 130 to 135 of H3 and 6 to 18 of H2B were found to be exposed, and interacted with antibodies speci®c for these domains, whereas the regions 1 to 11, 26 to 35 and 36 to 43 of H2B, and 80 to 89 and 85 to 102 of H4 were not accessible. Changes in the surface topography of nucleosomes induced by H4 acetylation, H1 and H3 phosphorylation, H1 and H2B poly(ADP-ribosyl)ation, H1 depletion and pH changes within a physiological range were characterized with antibody probes (Huletsky et al, 1989;Muller et al, 1982Muller et al, , 1984Thibeault et al, 1992). However, a limited number of polyclonal antibodies to synthetic peptides and monoclonal antibodies to histones were available at the time of these earlier studies and therefore only a few histone regions could be followed.…”

Section: Introductionmentioning

confidence: 98%

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

Stemmer

Briand

Muller

1997

Journal of Molecular Biology

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Section: Reactivity Of Histone Antibodies With Nucleosomes In a Direcsupporting

confidence: 94%

Section: Introductionmentioning

confidence: 98%

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

Stemmer

Briand

Muller

1997

Journal of Molecular Biology

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“…The epitope in the region H2B 36 -50 is, however, accessible to antibody binding [26] and is evidently not involved in hlstone-histone or histone-DNA interactions. The epitopes in H4 80-89 and 85-102 are inaccessible to antibody [28], consistent with protection of the lysine-containing peptide H4 79-92 by DNA. The C-terminal hexapeptide of H3 is accessible to antibodies both in chromatin 1281 and in nucleosome core particles [27].…”

Section: Discussionsupporting

confidence: 60%

“…6. Antibodies raised against particular peptide epitopes and tested for binding to chromatin showed that epitope(s) in the region H2B 6 -18 were accessible to antibody whereas those in H2B 26-35 and H2B 36-43 were not [28]. The most likely explanation, in view of the protection against chemical modification afforded by DNA to Lys-30 and Lys-31, is that the region 26 -43 of H2B interacts extensively with DNA.…”

Section: Discussionmentioning

confidence: 99%

Lysine‐containing DNA‐binding regions on the surface of the histone octamer in the nucleosome core particle

Lambert

Thomas

1986

European Journal of Biochemistry

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The DNA bound on the surface of the histone octamer in the nucleosome core particle partially protects the &-amino side-chains of a subset of the lysine residues from reductive methylation. Most of the strongly protected lysines, which probably define the path of the DNA on the octamer surface, are in the globular ('structured') regions of the core histones rather than in the N-terminal or C-terminal 'tails'. Analysis of the protected peptides shows that the three strongest lysine-containing DNA-binding sites in the core histones contain the sequence -Lys/ Arg-Lys-Thr/Ser-. On the assumption that the lysine-containing regions protected from chemical modification are also those found in lysine-DNA cross-links in another study [Mirzabekov et al. (1978) Proc. Nut1 Acad. Sci. USA 75, 41 84 -41 881, particular DNA-protected peptides may be tentatively assigned to particular DNA contact points. T h s leads to a more detailed description of the DNA-binding regions on the octamer surface in the nucleosome core particle. Strong contacts, reflected in strongly protected lysines, may well contribute to the distortion of the DNA from smooth bending [Richmond et al. (1984) Nature (Lond.) 311, 532-5371.A low-resolution (2.0 nm) structure, deduced for the nucleosome core particle, defined in outline the region within the structure occupied by particular histones, and the path of the DNA [1]. Subsequently these features were determined directly from X-ray diffraction analysis of nucleosome core particle crystals at 0.7 nm resolution [2]. However, the details of histone chain folding and of histone-DNA interactions remain at present unresolved. Some interactions may be particularly strong and may be responsible for distortion of the DNA from otherwise smooth bending [2]. They seem likely to involve electrostatic linkages between lysine and arginine side-chains on the surface of the histone octamer and DNA phosphates.We describe here a procedure for specifically radiolabelling lysine residues that are afforded protection against chemical modification in the native structure by the DNA. These are presumed to be engaged in electrostatic interaction with DNA phosphates and to direct the path of the DNA on the octarner surface. MATERIALS AND METHODSPreparation of nucleosome core particles and 'tail-less' core particles Chromatin was prepared from chicken erythrocyte nuclei [4] at AZ60 = 100 (read in 1 M NaOH) by digestion with micrococcal nuclease (5 units/ml nuclei for 4.25 min at 37°C) NazEDTA, pH 7.5. H1 and H5 were removed by treatment with the cation-exchange resin, AG5OW-X2 [6] and the stripped chromatin was dialysed into 10 mM Tris/HCl, 0.1 mM Na2EDTA, 0.25 mM phenylmethylsulphonyl fluoride (PMSF), pH 7.5, concentrated against solid sucrose and redialysed against the same buffer. It was digested to 146-basepair nucleosome core particles with micrococcal nuclease, under conditions determined from an analytical time course of digestion and analysis of the extracted DNA in a 6% polyacrylamide/7 M urea gel. The bulk digest wa...

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“…Using the two large cleavage H2B fragments 1-59 and 63-125 and sera from lupus patients, Hardin and Thomas also drew the same conclusion (22). Interestingly in this context, it has also been shown that the N-terminal region of H2B is readily exposed at the surface of mono-and polynucleosomes (29,38,39). The predominant reactivity toward this region of the core histone H2B is very likely to be directly correlated to its structural localization in the nucleosomal particle.…”

Section: Discussionmentioning

confidence: 88%

Identification of New Pathogenic Players in Lupus: Autoantibody-Secreting Cells Are Present in Nephritic Kidneys of (NZBxNZW)F1 Mice

Lacotte

Dumortier

Décossas

et al. 2010

The Journal of Immunology

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An important hallmark of systemic lupus erythematosus is the production of autoantibodies specific for nuclear Ags, among which nucleosomes and their constituents, DNA and histones. It is widely admitted that some of these autoantibodies contribute largely in lupus pathogenesis because of their nephritogenic potential. However, the underlying mechanisms are still debated. In this study, we analyzed the autoimmune response against histone H2B during the course of the disease in lupus-prone (NZBxNZW)F1 mice, both in lymphoid organs and kidneys, and we assessed its potential involvement in lupus pathogenicity. We found that the N-terminal region of histone H2B represents a preferential target for circulating autoantibodies, which kinetics of appearance positively correlates with disease development. Furthermore, immunization of preautoimmune (NZBxNZW)F1 mice with H2B peptide 1–25 accelerates the disease. Kidney eluates from diseased (NZBxNZW)F1 mice do contain IgG Abs reacting with this peptide, and this H2B sequence was found to be accessible to specific Ab probes in Ag-containing deposits detected in nephritic kidneys. Finally, compared with control normal mice and to young preautoimmune (NZBxNZW)F1 animals, the frequency of cells secreting autoantibodies reacting with peptide 1–25 was significantly raised in the spleen and bone marrow and most importantly on a pathophysiological point of view, locally, in nephritic kidneys of diseased (NZBxNZW)F1 mice. Altogether our results demonstrate the existence in (NZBxNZW)F1 mice of both a systemic and local B cell response targeting the N-terminal region of histone H2B, and highlight the potential implication of this nuclear domain in lupus pathology.

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Use of histone antibodies for studying chromatin topography and the phosphorylation of chromatin subunits.

Cited by 47 publications

References 62 publications

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

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

Lysine‐containing DNA‐binding regions on the surface of the histone octamer in the nucleosome core particle

Identification of New Pathogenic Players in Lupus: Autoantibody-Secreting Cells Are Present in Nephritic Kidneys of (NZBxNZW)F1 Mice

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