TY  - JOUR
AU  - Royet, Adrien
AU  - Ruedas, Rémi
AU  - Gargowitsch, Laetitia
AU  - Gervais, Virginie
AU  - Habersetzer, Johann
AU  - Pieri, Laura
AU  - Ouldali, Malika
AU  - Paternostre, Maïté
AU  - Hofmann, Ilse
AU  - Tubiana, Thibault
AU  - Fieulaine, Sonia
AU  - Bressanelli, Stéphane
TI  - Nonstructural protein 4 of human norovirus self-assembles into various membrane-bridging multimers.
JO  - The journal of biological chemistry
VL  - 300
SN  - 0021-9258
CY  - Bethesda, Md.
PB  - Soc.
M1  - DKFZ-2024-01770
SP  -  107724
PY  - 2024
N1  - Volume 300, Issue 9, September 2024, 107724 / Core Facility Antibodies
AB  - Single-stranded, positive-sense RNA ((+)RNA) viruses replicate their genomes in virus-induced intracellular membrane compartments. (+)RNA viruses dedicate a significant part of their small genomes (a few thousands to a few tens of thousands of bases) to the generation of these compartments by encoding membrane-interacting proteins and/or protein domains. Noroviruses are a very diverse genus of (+)RNA viruses including human and animal pathogens. Human noroviruses are the major cause of acute gastroenteritis worldwide, with genogroup II genotype 4 (GII.4) noroviruses accounting for the vast majority of infections. Three viral proteins encoded in the N-terminus of the viral replication polyprotein direct intracellular membrane rearrangements associated with norovirus replication. Of these three, nonstructural protein 4 (NS4) seems to be the most important, although its exact functions in replication organelle formation are unknown. Here we produce, purify and characterize GII.4 NS4. AlphaFold modeling combined with experimental data refine and correct our previous crude structural model of NS4. Using simple artificial liposomes, we report an extensive characterization of the membrane properties of NS4. We find that NS4 self-assembles and thereby bridges liposomes together. Cryo-EM, NMR and membrane flotation show formation of several distinct NS4 assemblies, at least two of them bridging pairs of membranes together in different fashions. Noroviruses belong to (+)RNA viruses whose replication compartment is extruded from the target endomembrane and generates double-membrane vesicles. Our data establish that the 21-kDa GII.4 human norovirus NS4 can, in the absence of any other factor, recapitulate in tubo several features, including membrane apposition, that occur in such processes.
KW  - Cryo-electron microscopy (Other)
KW  - Lipid-protein interaction (Other)
KW  - Liposome (Other)
KW  - Membrane (Other)
KW  - Nonstructural protein (Other)
KW  - Norovirus (Other)
KW  - Plus-stranded RNA virus (Other)
KW  - Protein assembly (Other)
KW  - Viral protein (Other)
KW  - Viral replication (Other)
LB  - PUB:(DE-HGF)16
C6  - pmid:39214299
DO  - DOI:10.1016/j.jbc.2024.107724
UR  - https://inrepo02.dkfz.de/record/292524
ER  -