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J Mol Biol ; 435(16): 168153, 2023 08 15.
Artículo en Inglés | MEDLINE | ID: mdl-37210029

RESUMEN

Viral factories of liquid-like nature serve as sites for transcription and replication in most viruses. The respiratory syncytial virus factories include replication proteins, brought together by the phosphoprotein (P) RNA polymerase cofactor, present across non-segmented negative stranded RNA viruses. Homotypic liquid-liquid phase separation of RSV-P is governed by an α-helical molten globule domain, and strongly self-downmodulated by adjacent sequences. Condensation of P with the nucleoprotein N is stoichiometrically tuned, defining aggregate-droplet and droplet-dissolution boundaries. Time course analysis show small N-P nuclei gradually coalescing into large granules in transfected cells. This behavior is recapitulated in infection, with small puncta evolving to large viral factories, strongly suggesting that P-N nucleation-condensation sequentially drives viral factories. Thus, the tendency of P to undergo phase separation is moderate and latent in the full-length protein but unleashed in the presence of N or when neighboring disordered sequences are deleted. This, together with its capacity to rescue nucleoprotein-RNA aggregates suggests a role as a "solvent-protein".


Asunto(s)
Nucleoproteínas , Virus Sincitial Respiratorio Humano , Compartimentos de Replicación Viral , Proteínas Estructurales Virales , ARN Polimerasas Dirigidas por ADN/metabolismo , Nucleoproteínas/metabolismo , Virus Sincitial Respiratorio Humano/metabolismo , Virus Sincitial Respiratorio Humano/fisiología , Compartimentos de Replicación Viral/metabolismo , Replicación Viral , Proteínas Estructurales Virales/metabolismo , Humanos
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