Journal Article

3.9 Å structure of the nucleosome core particle determined by phase-plate cryo-EM

Eugene Y.D. Chua, Vinod K. Vogirala, Oviya Inian, Andrew S.W. Wong, Lars Nordenskiöld, Juergen M. Plitzko, Radostin Danev and Sara Sandin

in Nucleic Acids Research

Volume 44, issue 17, pages 8013-8019
Published in print September 2016 | ISSN: 0305-1048
Published online August 2016 | e-ISSN: 1362-4962 | DOI: http://dx.doi.org/10.1093/nar/gkw708

More Like This

Show all results sharing these subjects:

  • Molecular and Cell Biology
  • Genetics and Genomics
  • Research Methods in Life Sciences
  • Proteins
  • Molecular Biology and Genetics

GO

Show Summary Details

Preview

The Volta phase plate is a recently developed electron cryo-microscopy (cryo-EM) device that enables contrast enhancement of biological samples. Here we have evaluated the potential of combining phase-plate imaging and single particle analysis to determine the structure of a small protein–DNA complex. To test the method, we made use of a 200 kDa Nucleosome Core Particle (NCP) reconstituted with 601 DNA for which a high-resolution X-ray crystal structure is known. We find that the phase plate provides a significant contrast enhancement that permits individual NCPs and DNA to be clearly identified in amorphous ice. The refined structure from 26,060 particles has an overall resolution of 3.9 Å and the density map exhibits structural features consistent with the estimated resolution, including clear density for amino acid side chains and DNA features such as the phosphate backbone. Our results demonstrate that phase-plate cryo-EM promises to become an important method to determine novel near-atomic resolution structures of small and challenging samples, such as nucleosomes in complex with nucleosome-binding factors.

Journal Article.  3716 words.  Illustrated.

Subjects: Molecular and Cell Biology ; Genetics and Genomics ; Research Methods in Life Sciences ; Proteins ; Molecular Biology and Genetics

Users without a subscription are not able to see the full content. Please, subscribe or login to access all content.