# New rhinovirus uncoating intermediate reveals how sodium versus potassium ions influence RNA release

## TL;DR

Researchers discovered a new rhinovirus uncoating stage called the 'E0 particle' and found that sodium and potassium ions affect how the virus releases its RNA.

## Contribution

The study identifies a novel uncoating intermediate (E0 particle) and reveals how sodium and potassium ions influence rhinovirus RNA release.

## Key findings

- E0 particles form in acidic K+ phosphate buffer and retain VP4 and a pocket factor.
- RNA compactness differs in K+ versus Na+ phosphate buffers, with RNA unfolding observed in Na+.
- Phosphotungstate penetration and cryo-EM imaging showed structural differences between A and E0 particles.

## Abstract

Electron microscopy (EM) of rhinovirus A2 (RV-A2) incubated in Na+ phosphate buffer (pH 7.6) for 12 h at 25 °C revealed partial fragmentation, whereas upon incubation in K+ phosphate buffer, RV-A2 appeared intact. In buffers adjusted to pH 5.8, these differences became more pronounced; acidic Na+ phosphate buffer promoted disintegration of the particles, whereas in acidic K+ phosphate buffer, the virus appeared like native. Incubation in the acidic buffers for one hour at 4 °C followed by neutralisation resulted in the respective formation of non-infectious A particles (in Na+) and a non-infectious novel uncoating intermediate (in K+), which we termed ‘E0 particle’. Negative staining EM revealed phosphotungstate penetration into A particles, but not into E0 particles. Cryo-EM image reconstruction of the E0 particle showed clear differences between A and E0 particles; like native virus, E0 contained VP4 and a pocket factor. Native RV-A2 RNA cores, obtained by gentle proteinase-K digestion in K+ and Na+ phosphate buffer, respectively, differed in accessibility of dsRNA regions, detected by PaSTRy. Variance in RNA compactness observed in K+ versus Na+ phosphate buffer was confirmed by rotary shadowing EM; in K+ phosphate buffer, the RNA remained condensed while, in Na+ phosphate buffer, distinct unfolding stages were apparent.

The online version contains supplementary material available at 10.1038/s41598-025-20627-0.