A conserved motif in Henipavirus P/V/W proteins drives the fibrillation of the W protein from Hendra virus

Protein Science, Год журнала: 2025, Номер 34(4)

Опубликована: Март 18, 2025

The Hendra (HeV) and Nipah (NiV) viruses are high-priority, biosafety level-4 pathogens that cause fatal neurological respiratory disease. Their P gene encodes not only the protein, an essential polymerase cofactor, but also virulence factors V W. We previously showed W protein of HeV (WHeV) forms amyloid-like fibrils one its subdomains, PNT3, fibrillates in isolation. However, fibrillation kinetics is much faster case full-length WHeV compared to suggesting another region contributes process. In this work, we identified spanning residues 2-110 (PNT1) as crucial implicated fibrillation. Through site-directed mutagenesis, combined with thioflavin T binding experiments negative-staining transmission electron microscopy, a predicted cryptic amyloidogenic (CAR) within PNT1 main driver deciphered underlying molecular mechanism. Using FTIR, enriched cross β-sheets. Sequence alignment revealed conservation CAR across Henipavirus genus enabled identification hitherto never reported pro-amyloidogenic motif. ability form was experimentally shown be common property shared by proteins. Overall, study sheds light on mechanisms calls for future studies aimed at exploring relevance newly motif valuable target antiviral approaches.

Язык: Английский

Interactions between the picornavirus 3C(D) main protease and RNA induce liquid-liquid phase separation

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown

Опубликована: Апрель 17, 2025

ABSTRACT The picornavirus 3CD protein is a precursor to the 3C main protease and 3D RNA-dependent RNA polymerase. In addition its functions in proteolytic processing of virus polyprotein cleavage key host factors, domain interacts with cis-acting replication elements (CREs) within viral genome regulate translation events. We investigated molecular determinants binding using wide range biophysical computational methods. These studies showed that binds broad spectrum oligonucleotides, displaying minimal dependence on sequence structure. However, they also uncovered novel aspect these interactions, is, 3C-RNA can induce liquid-liquid phase separation (LLPS), 3CD-RNA interactions likewise leading LLPS. This may be general phenomenon for other 3C-like proteases, incorporating domains. findings have potential implications understanding virally induced apoptosis controlling stress granules, which involve LLPS include proteins known 3C/3CD.

Язык: Английский

Liquid-liquid phase separation in viral infection: from the occurrence and function to treatment potentials

Colloids and Surfaces B Biointerfaces, Год журнала: 2024, Номер 246, С. 114385 - 114385

Опубликована: Ноя. 17, 2024

Язык: Английский