Skip to content Skip to navigation

Visualizing the global secondary structure of a viral RNA genome with cryo-electron microscopy

TitleVisualizing the global secondary structure of a viral RNA genome with cryo-electron microscopy
Publication TypeJournal Article
Year of Publication2015
AuthorsGarmann, RF, Gopal, A, Athavale, SS, Knobler, CM, Gelbart, WM, Harvey, SC
Journalrna
Volume21
Pagination877–886
Abstract

The lifecycle, and therefore the virulence, of single-stranded (ss)-RNA viruses is regulated not only by their particular protein gene products, but also by the secondary and tertiary structure of their genomes. The secondary structure of the entire genomic RNA of satellite tobacco mosaic virus (STMV) was recently determined by selective 2′-hydroxyl acylation analyzed by primer extension (SHAPE). The SHAPE analysis suggested a single highly extended secondary structure with much less branching than occurs in the ensemble of structures predicted by purely thermodynamic algorithms. Here we examine the solution-equilibrated STMV genome by direct visualization with cryo-electron microscopy (cryo-EM), using an RNA of similar length transcribed from the yeast genome as a control. The cryo-EM data reveal an ensemble of branching patterns that are collectively consistent with the SHAPE-derived secondary structure model. Thus, our results both elucidate the statistical nature of the secondary structure of large ss-RNAs and give visual support for modern RNA structure determination methods. Additionally, this work introduces cryo-EM as a means to distinguish between competing secondary structure models if the models differ significantly in terms of the number and/or length of branches. Furthermore, with the latest advances in cryo-EM technology, we suggest the possibility of developing methods that incorporate restraints from cryo-EM into the next generation of algorithms for the determination of RNA secondary and tertiary structures.

URLhttp://rnajournal.cshlp.org/content/21/5/877.short
DOI10.1261/rna.047506.114
research thumbnail: