Functionalized graphene-oxide grids enable high-resolution cryo-EM structures of the SNF2h-nucleosome complex without crosslinking

Un Seng Chio, Eugene Palovcak, Anton A. A. Smith, Henriette Autzen, Elise N. Muñoz, Zanlin Yu, Feng Wang, David A. Agard, Jean-Paul Armache (), Geeta J. Narlikar () and Yifan Cheng ()
Additional contact information
Un Seng Chio: University of California San Francisco
Eugene Palovcak: University of California San Francisco
Anton A. A. Smith: Stanford University
Henriette Autzen: University of California San Francisco
Elise N. Muñoz: University of California, San Francisco
Zanlin Yu: University of California San Francisco
Feng Wang: University of California San Francisco
David A. Agard: University of California San Francisco
Jean-Paul Armache: Pennsylvania State University
Geeta J. Narlikar: University of California San Francisco
Yifan Cheng: University of California San Francisco

Nature Communications, 2024, vol. 15, issue 1, 1-12

Abstract: Abstract Single-particle cryo-EM is widely used to determine enzyme-nucleosome complex structures. However, cryo-EM sample preparation remains challenging and inconsistent due to complex denaturation at the air-water interface (AWI). Here, to address this issue, we develop graphene-oxide-coated EM grids functionalized with either single-stranded DNA (ssDNA) or thiol-poly(acrylic acid-co-styrene) (TAASTY) co-polymer. These grids protect complexes between the chromatin remodeler SNF2h and nucleosomes from the AWI and facilitate collection of high-quality micrographs of intact SNF2h-nucleosome complexes in the absence of crosslinking. The data yields maps ranging from 2.3 to 3 Å in resolution. 3D variability analysis reveals nucleotide-state linked conformational changes in SNF2h bound to a nucleosome. In addition, the analysis provides structural evidence for asymmetric coordination between two SNF2h protomers acting on the same nucleosome. We envision these grids will enable similar detailed structural analyses for other enzyme-nucleosome complexes and possibly other protein-nucleic acid complexes in general.

Date: 2024
References: View references in EconPapers View complete reference list from CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41467-024-46178-y Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-46178-y

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-024-46178-y

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().