Nucleosome core particle stability and conformational change

Ausió, Juan; Seger, Dalia; Eisenberg, Henryk

doi:10.1016/0022-2836(84)90383-8

Cited by 127 publications

(99 citation statements)

References 45 publications

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“…The stoichiometry of the histone octamer to the DNA in the reconstituted 208-12 oligonucleosome complexes was determined as described previously (28). The reconstituted chromatin particles thus obtained (at a concentration of ϳ40 -50 g/ml) were dialyzed against the appropriate buffers (25, 31) and used for subsequent analytical ultracentrifuge analysis (32,33). In some instances, the reconstituted nucleosome particles were concentrated 5-fold at 4°C using Centricon YM-10 or YM-50 (Millipore Corp., Bedford, MA).…”

Section: Methodsmentioning

confidence: 99%

Characterization of the Stability and Folding of H2A.Z Chromatin Particles

Abbott

Ivanova

Wang

et al. 2001

Journal of Biological Chemistry

146

110

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Section: Methodsmentioning

confidence: 99%

Characterization of the Stability and Folding of H2A.Z Chromatin Particles

Abbott

Ivanova

Wang

et al. 2001

Journal of Biological Chemistry

146

110

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“…The conformation of nucleosomes is known to be sensitive to the ionic concentration of the medium (Ausio et al, 1984;Yager & van Holde, 1984), and ionic concentration is one of the decisive factors in the polarity of the medium. Therefore, for this study, a sulfhydryl-specific fluorescence probe, 1,5-IAEDANS, was selected as the polarity probe (Hudson & Weber, 1973).…”

Section: ~~mentioning

confidence: 99%

Dynamics of interaction of RNA polymerase II with nucleosomes. I. Effect of salts

Bhargava

1993

Protein Science

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Mononucleosomes were labeled with the sulfhydryl-specific fluorescence probe 1,5-IAEDANS (5-(2-((iodoacety1)amino)ethyl)amino-naphthalene-1-sulfonic acid) by attaching the dye to the single cysteine of H3 through a covalent linkage. The enzyme RNA polymerase I1 (pol 11) utilized the native and the reconstituted fluorescent nucleosomes as templates with greatest efficiency when 0.2 M potassium acetate (AcOK) was used as the supporting salt; 0.2 M NaCl was found to be very much inhibitory. Measurement of polarity of the microenvironment of the dye at its binding site in the nucleosome showed the conformation to be more open in the presence of AcOK, compared to that in 0.1 or 0.2 M NaCl. The binding of pol I1 to the nucleosome resulted in a relatively more compact structure when measured in terms of the polarity of the microenvironment of the dye in various salt-dependent conformations of the nucleosomes. Time-resolved fluorescence spectroscopy showed that the probe molecule at its binding site undergoes certain excited-state processes, and the presence/absence or rate of these excited-state processes depends on the conformation of nucleosomes, which in turn depends on the type and concentration of the ion present in the medium. Time-resolved emission spectra showed that binding of nucleosomes by pol 11 established some new contacts that resulted in inaccessibility of the dye to the bulk solvent, reflecting a more hydrophobic environment for the dye in the steady-state spectra. Thus, binding or transcription of nucleosomes by pol I1 did not break open their structure. Rather, some transient internal adjustments within the histone octamer may take place to accommodate the bulky pol I1 molecule.

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Section: Nucleosome Core Particle Reconstitution Upon Renaturation Ofmentioning

confidence: 99%

“…Fraction C shows the presence of two main populations of macromolecules sedimenting at 10.6p0.2 (40 %) and 5.4p0.2 S (40 %), the latter one corresponding to free 145-bp DNA [26]. In this fraction, as well as in fractions B and D, there is a small amount (5-15 %) of material sedimenting with S 11, which reflects the binding of extra histones to the nucleosome core particle [26], which is most probably due to an error in the estimation of the histone\DNA ratio. Fractions A, B and D also show a variable amount (10-15 %) of 11 S sedimenting material, which results from the partial dissociation of the nucleosome core particles, which is a result of the experimental conditions of ionic strength and temperature [26].…”

Section: Nucleosome Core Particle Reconstitution Upon Renaturation Ofmentioning

confidence: 99%

“…The fraction sedimenting as 5.7 S corresponds to naked DNA, as corroborated by native PAGE (results not shown). At this point it is not clear if this behaviour is inherent to the intrinsic properties of the lamprey histones which lead to nucleosomes with a slightly decreased stability under the ionic conditions at which the samples were analysed (100 mM NaCl) [26], or to problems that could have arisen during the reconstitution procedure or handling of the sample before its analysis. Figure 1) For the CD analysis, the samples were dialysed against 10 mM sodium phosphate buffer, pH 7.2.…”

Section: Figure 4 Dnase I Footprinting Analysis Of Native Nucleosome mentioning

confidence: 99%

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Reconstitution of native-like nucleosome core particles from reversed-phase-HPLC-fractionated histones

Moore

Rice

Maya

et al. 1997

Biochemical Journal

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We have reconstituted nucleosome core particles from reversed-phase-HPLC-purified chicken erythrocyte core histones and 145 bp random-sequence DNA fragments. Characterization of the resulting nucleoprotein complexes by sedimentation velocity, CD and DNase I footprinting showed that they are structurally indistinguishable from native nucleosome core particles. Furthermore, we have shown that the ability to reproduce these native-like structural features in these reconstituted nucleosome core particles is basically independent of the biological source or the method used (i.e. salt versus acid) for the extraction of histones before their HPLC fractionation. The usefulness and relevance of this approach for the reconstitution of native-like chromatin structures from histone types (histone variants/post-translationally modified histones), which are usually available only in relatively small amounts, is discussed.

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Nucleosome core particle stability and conformational change

Cited by 127 publications

References 45 publications

Characterization of the Stability and Folding of H2A.Z Chromatin Particles

Characterization of the Stability and Folding of H2A.Z Chromatin Particles

Dynamics of interaction of RNA polymerase II with nucleosomes. I. Effect of salts

Reconstitution of native-like nucleosome core particles from reversed-phase-HPLC-fractionated histones

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