Self-Assembly of Measles Virus Nucleocapsid-like Particles: Kinetics and RNA Sequence-Dependence.

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Milles, Sigrid | Jensen, Malene Ringkjøbing | Communie, Guillaume | Maurin, Damien | Schoehn, Guy | Ruigrok, Rob W H | Blackledge, Martin

Edité par CCSD ; Wiley-VCH Verlag -

International audience. Measles virus RNA genomes are packaged into helical nucleocapsids (NCs), comprising thousands of nucleo-proteins (N) that bind the entire genome. N-RNA provides the template for replication and transcription by the viral polymerase and is a promising target for viral inhibition. Elucidation of mechanisms regulating this process has been severely hampered by the inability to controllably assemble NCs. Here, we demonstrate self-organization of N into NC-like particles in vitro upon addition of RNA, providing a simple and versatile tool for investigating assembly. Real-time NMR and fluorescence spectroscopy reveals biphasic assembly kinetics. Remarkably, assembly depends strongly on the RNA-sequence, with the genomic 5' end and poly-Adenine sequences assembling efficiently, while sequences such as poly-Uracil are incompetent for NC formation. This observation has important consequences for understanding the assembly process.

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