Structure determination of a DNA crystal by MicroED

Alison Haymaker; Andrey A Bardin; Tamir Gonen; Michael W Martynowycz; Brent L Nannenga

doi:10.1016/j.str.2023.07.005

Structure determination of a DNA crystal by MicroED

Structure. 2023 Dec 7;31(12):1499-1503.e2. doi: 10.1016/j.str.2023.07.005. Epub 2023 Aug 3.

Authors

Alison Haymaker¹, Andrey A Bardin¹, Tamir Gonen², Michael W Martynowycz³, Brent L Nannenga⁴

Affiliations

¹ Biodesign Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA; School for Engineering of Matter, Transport and Energy, Arizona State University, Tempe, AZ, USA.
² Department of Biological Chemistry, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Physiology, University of California, Los Angeles, Los Angeles, CA 90095, USA; Howard Hughes Medical Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA.
³ Department of Biological Chemistry, University of California, Los Angeles, Los Angeles, CA 90095, USA. Electronic address: mikewm@g.ucla.edu.
⁴ Biodesign Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, 727 East Tyler Street, Tempe, AZ 85287, USA; School for Engineering of Matter, Transport and Energy, Arizona State University, Tempe, AZ, USA. Electronic address: Brent.Nannenga@asu.edu.

PMID: 37541248
PMCID: PMC10805983 (available on 2024-12-07)
DOI: 10.1016/j.str.2023.07.005

Abstract

Microcrystal electron diffraction (MicroED) is a powerful tool for determining high-resolution structures of microcrystals from a diverse array of biomolecular, chemical, and material samples. In this study, we apply MicroED to DNA crystals, which have not been previously analyzed using this technique. We utilized the d(CGCGCG)₂ DNA duplex as a model sample and employed cryo-FIB milling to create thin lamella for diffraction data collection. The MicroED data collection and subsequent processing resulted in a 1.10 Å resolution structure of the d(CGCGCG)₂ DNA, demonstrating the successful application of cryo-FIB milling and MicroED to the investigation of nucleic acid crystals.

Keywords: DNA; cryo-electron microscopy; cryo-focused ion beam milling; microcrystal electron diffraction, MicroED; nucleic acids.

MeSH terms

Cryoelectron Microscopy / methods
Electrons*

Abstract

MeSH terms

Grants and funding