Self-Assembly of Measles Virus Nucleocapsid-like Particles: Kinetics and RNA Sequence Dependence.
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.
Univ. Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale, Grenoble, France.
Sigrid
S
Milles
Univ. Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale, Grenoble, France.
Malene Ringkjøbing
MR
Jensen
Univ. Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale, Grenoble, France.
Guillaume
G
Communie
Univ. Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale, Grenoble, France.
Damien
D
Maurin
Univ. Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale, Grenoble, France.
Guy
G
Schoehn
Univ. Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale, Grenoble, France. rob.ruigrok@ibs.fr.
Rob W H
RW
Ruigrok
Univ. Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale, Grenoble, France. martin.blackledge@ibs.fr.
Martin
M
Blackledge
D001483:Base Sequence; D007700:Kinetics; D008459:Measles virus / Q000378:metabolism*; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D019251:Nucleocapsid / Q000737:chemistry / Q000378:metabolism*; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000737:chemistry / Q000378:metabolism*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism*; D019065:Virus Assembly
D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D012367:RNA, Viral; D014764:Viral Proteins; C058314:nucleoprotein, Measles virus
NMR spectroscopy; fluorescence spectroscopy; measles; nucleocapsids; self-assembly