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Angew Chem Int Ed Engl. 2016 Aug 1;55(32):9356-60. doi: 10.1002/anie.201602619. Epub 2016 Jun 6.

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

Author information

1
Univ. Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale, Grenoble, France.
2
Univ. Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale, Grenoble, France. rob.ruigrok@ibs.fr.
3
Univ. Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale, Grenoble, France. martin.blackledge@ibs.fr.

Abstract

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.

KEYWORDS:

NMR spectroscopy; fluorescence spectroscopy; measles; nucleocapsids; self-assembly

PMID:
27270664
PMCID:
PMC6680290
DOI:
10.1002/anie.201602619
[Indexed for MEDLINE]
Free PMC Article

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