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Radiation-damage investigation of a DNA 16-mer

DOI: 10.1107/S160057751900763X DOI Help

Authors: Valeria Bugris (Biological Research Centre, HAC-BRC; University of Oxford) , Veronika Harmat (Biological Research Centre, HAC-BRC; Eötvös Loránd University) , Györgyi Ferenc (Biological Research Centre, HAC-BRC) , Sándor Brockhauser (Biological Research Centre, HAC-BRC; European X-ray Free-Electron Laser Facility GmbH (EuXFEL)) , Ian Carmichael (University of Notre Dame) , Elspeth F. Garman (University of Oxford)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of Synchrotron Radiation , VOL 26

State: Published (Approved)
Published: July 2019
Diamond Proposal Number(s): 18069

Abstract: In macromolecular crystallography, a great deal of effort has been invested in understanding radiation-damage progression. While the sensitivity of protein crystals has been well characterized, crystals of DNA and of DNA–protein complexes have not thus far been studied as thoroughly. Here, a systematic investigation of radiation damage to a crystal of a DNA 16-mer diffracting to 1.8 Å resolution and held at 100 K, up to an absorbed dose of 45 MGy, is reported. The RIDL (Radiation-Induced Density Loss) automated computational tool was used for electron-density analysis. Both the global and specific damage to the DNA crystal as a function of dose were monitored, following careful calibration of the X-ray flux and beam profile. The DNA crystal was found to be fairly radiation insensitive to both global and specific damage, with half of the initial diffraction intensity being lost at an absorbed average diffraction-weighted dose, D1/2, of 19 MGy, compared with 9 MGy for chicken egg-white lysozyme crystals under the same beam conditions but at the higher resolution of 1.4 Å. The coefficient of sensitivity of the DNA crystal was 0.014 Å2 MGy−1, which is similar to that observed for proteins. These results imply that the significantly greater radiation hardness of DNA and RNA compared with protein observed in a DNA–protein complex and an RNA–protein complex could be due to scavenging action by the protein, thereby protecting the DNA and RNA in these studies. In terms of specific damage, the regions of DNA that were found to be sensitive were those associated with some of the bound calcium ions sequestered from the crystallization buffer. In contrast, moieties farther from these sites showed only small changes even at higher doses.

Journal Keywords: DNA; radiation damage; global damage; specific damage

Subject Areas: Biology and Bio-materials

Instruments: I24-Microfocus Macromolecular Crystallography

Added On: 27/06/2019 09:07

Discipline Tags:

Life Sciences & Biotech Structural biology

Technical Tags:

Diffraction Macromolecular Crystallography (MX)