title: Feline Calicivirus Infection Disrupts Assembly of Cytoplasmic Stress Granules and Induces G3BP1 Cleavage.
abstract: In response to stress such as virus infection, cells can stall translation by storing mRNAs away in cellular compartments called stress granules (SGs). This defense mechanism favors cell survival by limiting the use of energy and nutrients until the stress is resolved. In some cases it may also block viral propagation as viruses are dependent on the host cell resources to produce viral proteins. Human norovirus is a member of the Caliciviridae family responsible for gastroenteritis outbreaks worldwide. Previous studies on caliciviruses have identified mechanisms by which they can usurp the host translational machinery, using the viral protein genome-linked VPg, or regulate host protein synthesis through the mitogen-activated protein kinase (MAPK) pathway. Here, we examined the effect of feline calicivirus (FCV) infection on SG accumulation. We show that FCV infection impairs the assembly of SGs despite an increased phosphorylation of eukaryotic initiation factor eIF2α, a hallmark of stress pathway activation. Furthermore, SGs did not accumulate in FCV-infected cells that were stressed with arsenite or hydrogen peroxide. FCV infection resulted in the cleavage of the SG-nucleating protein Ras-GTPase activating SH3 domain-binding protein (G3BP1), which is mediated by the viral 3C-like proteinase NS6(Pro) Using mutational analysis, we identified the FCV-induced cleavage site within G3BP1, which differs from the poliovirus 3C proteinase cleavage site previously identified. Finally, we showed that NS6(Pro)-mediated G3BP1 cleavage impairs SG assembly. In contrast, murine norovirus (MNV) infection did not impact arsenite-induced SG assembly or G3BP1 integrity, suggesting that related caliciviruses have distinct effects on the stress response pathway.
  
  
  
  Human noroviruses are a major cause of viral gastroenteritis, and it is important to understand how they interact with the infected host cell. Feline calicivirus (FCV) and murine norovirus (MNV) are used as models to understand norovirus biology. Recent studies have suggested that the assembly of stress granules is central in orchestrating stress and antiviral responses to restrict viral replication. Overall, our study provides the first insight on how caliciviruses impair stress granule assembly by targeting the nucleating factor G3BP1 via the viral proteinase NS6(Pro) This work provides new insights into host-pathogen interactions that regulate stress pathways during FCV infection.
authors.affiliation: University of Surrey, Faculty of Health and Medical Sciences, School of Biosciences and Medicine, Guildford, United Kingdom.
authors.forename: Majid N
authors.initials: MN
authors.lastname: Humoud
authors.affiliation: University of Surrey, Faculty of Health and Medical Sciences, School of Biosciences and Medicine, Guildford, United Kingdom.
authors.forename: Nicole
authors.initials: N
authors.lastname: Doyle
authors.affiliation: University of Surrey, Faculty of Health and Medical Sciences, School of Biosciences and Medicine, Guildford, United Kingdom.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Royall
authors.affiliation: University of Surrey, Faculty of Health and Medical Sciences, School of Biosciences and Medicine, Guildford, United Kingdom.
authors.forename: Margaret M
authors.initials: MM
authors.lastname: Willcocks
authors.affiliation: Division of Virology, Department of Pathology, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge, United Kingdom.
authors.forename: Frederic
authors.initials: F
authors.lastname: Sorgeloos
authors.affiliation: Virology Division, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Frank
authors.initials: F
authors.lastname: van Kuppeveld
authors.affiliation: University of Surrey, Faculty of Health and Medical Sciences, School of Biosciences and Medicine, Guildford, United Kingdom.
authors.forename: Lisa O
authors.initials: LO
authors.lastname: Roberts
authors.affiliation: Division of Virology, Department of Pathology, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge, United Kingdom.
authors.forename: Ian G
authors.initials: IG
authors.lastname: Goodfellow
authors.affiliation: Virology Division, Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Martijn A
authors.initials: MA
authors.lastname: Langereis
authors.affiliation: University of Surrey, Faculty of Health and Medical Sciences, School of Biosciences and Medicine, Guildford, United Kingdom n.locker@surrey.ac.uk.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Locker
mesh_terms: D000086042:3C Viral Proteases; D000818:Animals; D017250:Caliciviridae Infections / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D017927:Calicivirus, Feline / Q000472:pathogenicity*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002415:Cats; D003546:Cysteine Endopeptidases / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000378:metabolism* / Q000821:virology; D004265:DNA Helicases; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D006367:HeLa Cells; D054884:Host-Pathogen Interactions; D006801:Humans; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D010766:Phosphorylation; D000075223:Poly-ADP-Ribose Binding Proteins; D020365:RNA Helicases; D000067776:RNA Recognition Motif Proteins; D014764:Viral Proteins / Q000378:metabolism; D014779:Virus Replication
chemical_list: D002352:Carrier Proteins; D015852:Eukaryotic Initiation Factor-2; D000075223:Poly-ADP-Ribose Binding Proteins; D000067776:RNA Recognition Motif Proteins; D014764:Viral Proteins; D020928:Mitogen-Activated Protein Kinases; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases; D004265:DNA Helicases; C116070:G3BP1 protein, human; D020365:RNA Helicases