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Structure Change from β-Strand and Turn to α-Helix in Histone H2A-H2B Induced by DNA Damage Response - PubMed

  • ️Fri Jan 01 2016

Structure Change from β-Strand and Turn to α-Helix in Histone H2A-H2B Induced by DNA Damage Response

Yudai Izumi et al. Biophys J. 2016.

Abstract

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.

Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

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Figures

Figure 1
Figure 1

(a) SDS-PAGE analysis of H2A-H2B extracted from core histones in unirradiated cells and 40 Gy irradiated cells after 30 min incubation. The lane labeled M refers to the standard molecular weight marker (XL-Ladder (low range); APRO Life Science Institute, Tokushima, Japan). The lanes labeled U and I refer to H2A-H2B extracted from unirradiated and irradiated cells, respectively. uH2A(Z) refers to monoubiquitinated H2A(Z). (b) Western blot analysis obtained using the H2AZ antibody.

Figure 2
Figure 2

(a–c) CD spectra of H2A-H2B extracted from core histones in unirradiated (Unirrad., black) and x-irradiated (Irrad., red) cells, measured at (a) 294, (b) 310, and (c) 330 K. In (a), a CD spectrum of H2A-H2B irradiated with 40 Gy x-rays after extraction from unirradiated cells (13) is also shown (x-irrad. H2A-H2B, blue). To see this figure in color, go online.

Figure 3
Figure 3

(a–d) Temperature dependence of the contents of (a) α-helix, (b) β-strand, (c) turn, and (d) other structures of H2A-H2B extracted from core histones in unirradiated (Unirrad., open circle) and x-irradiated (Irrad., solid circle) cells. The contents of x-irradiated H2A-H2B at 294 K (X-irrad. H2A-H2B, solid triangle) are also shown.

Figure 4
Figure 4

(a) Temperature dependence of the CD intensity at 222 nm of H2A-H2B extracted from core histones in unirradiated (Unirrad., open circle) and x-irradiated (Irrad., solid circle) cells. Inset: magnification of the unirradiated sample between 320 and 334 K. (b) Temperature dependence of ΔG of both samples, and the corresponding curves of the best fit obtained by the weighted least squares method.

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