Phosphorylation at the homotypic interface regulates nucleoprotein oligomerization and assembly of the influenza virus replication machinery

Negative-sense RNA viruses assemble large ribonucleoprotein (RNP) complexes that direct replication and transcription of the viral genome. Influenza virus RNPs contain the polymerase, genomic RNA and multiple copies of nucleoprotein (NP). During RNP assembly, monomeric NP oligomerizes along the leng...

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Bibliographic Details
Published in:PLoS pathogens 2015-04, Vol.11 (4), p.e1004826-e1004826
Main Authors: Mondal, Arindam, Potts, Gregory K, Dawson, Anthony R, Coon, Joshua J, Mehle, Andrew
Format: Article
Language:English
Subjects:
Animals
Dogs
Experiments
Gene expression
Genetic aspects
Genomes
Health aspects
HEK293 Cells
Humans
Identification and classification
Immunoprecipitation
Infections
Influenza
Influenza A virus - physiology
Influenza B virus - physiology
Influenza viruses
Influenza, Human - metabolism
Kinases
Machinery
Madin Darby Canine Kidney Cells
Mass Spectrometry
Nucleoproteins - metabolism
Peptides
Phosphorylation
Proteins
Scientific imaging
Virus replication
Virus Replication - physiology
Viruses
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Summary:Negative-sense RNA viruses assemble large ribonucleoprotein (RNP) complexes that direct replication and transcription of the viral genome. Influenza virus RNPs contain the polymerase, genomic RNA and multiple copies of nucleoprotein (NP). During RNP assembly, monomeric NP oligomerizes along the length of the genomic RNA. Regulated assembly of the RNP is essential for virus replication, but how NP is maintained as a monomer that subsequently oligomerizes to form RNPs is poorly understood. Here we elucidate a mechanism whereby NP phosphorylation regulates oligomerization. We identified new evolutionarily conserved phosphorylation sites on NP and demonstrated that phosphorylation of NP decreased formation of higher-order complexes. Two phosphorylation sites were located on opposite sides of the NP:NP interface. In both influenza A and B virus, mutating or mimicking phosphorylation at these residues blocked homotypic interactions and drove NP towards a monomeric form. Highlighting the central role of this process during infection, these mutations impaired RNP formation, polymerase activity and virus replication. Thus, dynamic phosphorylation of NP regulates RNP assembly and modulates progression through the viral life cycle.
ISSN:1553-7374
1553-7366
1553-7374
DOI:10.1371/journal.ppat.1004826