We report the global structural alterations in histone H3 proteins induced by lysine-9 mono-, di- and trimethylation, which are part of the critical post-translational modifications for DNA damage responses, identified using synchrotron radiation circular dichroism (CD) spectroscopy. Compared with unmodified H3, mono- and dimethylation increases the number of α-helices and decreases the numbers of β-strands, while trimethylation decreases the α-helix content and increases the β-strand content. Comparison of the secondary-structure contents of these histone H3 proteins suggests that the methylation-induced structural alterations occur at residues not only close to but also distant from the methylated sites. Such global structural alterations may regulate the interactions of methylated histones with other molecules, such as histone-binding proteins in DNA damage repair processes.

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Using synchrotron radiation-based circular dichroism spectroscopy, we found that the DNA damage response induces an increase of α-helix structure and a decrease of β-strand and turn structures in histone H2A-H2B extracted from x-irradiated human HeLa cells. The structural alterations correspond to the assumption that an average of eight amino acid residues form new α-helix structures at 310 K. We propose the structural transition from β-strand and turn structures to an α-helix structure in H2A-H2B as a novel, to our knowledge, process involved in the DNA damage response.

Deployments of spherical grids to obtain high energy and angular resolutions for retarding field analyzers (RFAs) having acceptance angles as large as or larger than ±45° were explored under the condition of using commercially available microchannel plates with effective diameters of approximately 100 mm. As a result of electron trajectory simulations, a deployment of three spherical grids with significantly different grid separations instead of conventional equidistant separations showed an energy resolving power (E/ΔE) of 3200 and an angular resolution of 0.6°. The mesh number of the wire mesh retarding grid used for the simulation was 250. An RFA constructed with the simulated design experimentally showed an E/ΔE of 1100 and an angular resolution of 1°. Using the RFA and synchrotron radiation of 900 eV, photoelectron diffraction (PED) measurements were performed for single-crystal graphite. A clear C 1s PED pattern was observed even when the differential energy of the RFA was set at 0.5 eV. Further improvement of the energy resolution was theoretically examined under the assumption of utilizing a retarding grid fabricated by making a large number of radially directed cylindrical holes through a partial spherical shell instead of using a wire mesh retarding grid. An E/ΔE of 14 500 was predicted for a hole design with a diameter of 60 μm and a depth of 100 μm. A retarding grid with this hole design and a holed area corresponding to an acceptance angle of ±7° was fabricated. An RFA constructed with this retarding grid experimentally showed an E/ΔE of 1800. Possible reasons for the experimental E/ΔE lower than the theoretical values are discussed.