Multiple capsid protein binding sites mediate selective packaging of the alphavirus genomic RNA
| Autor: | Markus Hafner, Dimitrios G. Anastasakis, Rebecca S. H. Brown, Margaret Kielian |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Models Molecular Science viruses Alphaviruses General Physics and Astronomy Viral transmission Alphavirus Semliki Forest virus Virus Replication Virus General Biochemistry Genetics and Molecular Biology Article 03 medical and health sciences Capsid Chlorocebus aethiops Animals Guide RNA Binding site lcsh:Science Nucleocapsid Vero Cells Multidisciplinary Binding Sites 030102 biochemistry & molecular biology biology Chemistry Virus Assembly Viral nucleocapsid RNA virus diseases General Chemistry Genomics biochemical phenomena metabolism and nutrition biology.organism_classification Semliki forest virus Cell biology 030104 developmental biology Viral replication Biotinylation RNA Viral lcsh:Q Capsid Proteins Chikungunya virus Protein Binding |
| Zdroj: | Nature Communications Nature Communications, Vol 11, Iss 1, Pp 1-16 (2020) |
| ISSN: | 2041-1723 |
| Popis: | The alphavirus capsid protein (Cp) selectively packages genomic RNA (gRNA) into the viral nucleocapsid to produce infectious virus. Using photoactivatable ribonucleoside crosslinking and an innovative biotinylated Cp retrieval method, here we comprehensively define binding sites for Semliki Forest virus (SFV) Cp on the gRNA. While data in infected cells demonstrate Cp binding to the proposed genome packaging signal (PS), mutagenesis experiments show that PS is not required for production of infectious SFV or Chikungunya virus. Instead, we identify multiple Cp binding sites that are enriched on gRNA-specific regions and promote infectious SFV production and gRNA packaging. Comparisons of binding sites in cytoplasmic vs. viral nucleocapsids demonstrate that budding causes discrete changes in Cp-gRNA interactions. Notably, Cp’s top binding site is maintained throughout virus assembly, and specifically binds and assembles with Cp into core-like particles in vitro. Together our data suggest a model for selective alphavirus genome recognition and assembly. Alphaviruses need to selectively package genomic viral RNA for transmission, but the packaging mechanism remains unclear. Here, Brown et al. combine PAR-CLIP with biotinylated capsid protein (Cp) retrieval and identify multiple Cp binding sites on genomic viral RNA that promote virion formation. |
| Databáze: | OpenAIRE |
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