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Activation of PI3K, Akt, and ERK during early rotavirus infection leads to V-ATPase-dependent endosomal acidification required for uncoating
- Source :
- PLoS Pathogens, PLoS Pathogens, Vol 14, Iss 1, p e1006820 (2018)
- Publication Year :
- 2018
- Publisher :
- Public Library of Science, 2018.
-
Abstract
- The cellular PI3K/Akt and/or MEK/ERK signaling pathways mediate the entry process or endosomal acidification during infection of many viruses. However, their roles in the early infection events of group A rotaviruses (RVAs) have remained elusive. Here, we show that late-penetration (L-P) human DS-1 and bovine NCDV RVA strains stimulate these signaling pathways very early in the infection. Inhibition of both signaling pathways significantly reduced production of viral progeny due to blockage of virus particles in the late endosome, indicating that neither of the two signaling pathways is involved in virus trafficking. However, immunoprecipitation assays using antibodies specific for pPI3K, pAkt, pERK and the subunit E of the V-ATPase co-immunoprecipitated the V-ATPase in complex with pPI3K, pAkt, and pERK. Moreover, Duolink proximity ligation assay revealed direct association of the subunit E of the V-ATPase with the molecules pPI3K, pAkt, and pERK, indicating that both signaling pathways are involved in V-ATPase-dependent endosomal acidification. Acidic replenishment of the medium restored uncoating of the RVA strains in cells pretreated with inhibitors specific for both signaling pathways, confirming the above results. Isolated components of the outer capsid proteins, expressed as VP4-VP8* and VP4-VP5* domains, and VP7, activated the PI3K/Akt and MEK/ERK pathways. Furthermore, psoralen-UV-inactivated RVA and CsCl-purified RVA triple-layered particles triggered activation of the PI3K/Akt and MEK/ERK pathways, confirming the above results. Our data demonstrate that multistep binding of outer capsid proteins of L-P RVA strains with cell surface receptors phosphorylates PI3K, Akt, and ERK, which in turn directly interact with the subunit E of the V-ATPase to acidify the late endosome for uncoating of RVAs. This study provides a better understanding of the RVA-host interaction during viral uncoating, which is of importance for the development of strategies aiming at controlling or preventing RVA infections.<br />Author summary Viral particles must transport their genome into the cytoplasm or the nucleus of host cells to initiate successful infection. Knowledge of how viruses may pirate host cell signaling cascades or molecules to promote their own replication can facilitate the development of antiviral drugs. Group A rotavirus (RVA) is a major etiological agent of acute gastroenteritis in young children and the young of various mammals. RVA enters cells by a complex multistep process. However, the cellular signaling cascades or molecules that facilitate these processes are incompletely understood. Here, we demonstrate that infection with late-penetration RVA strains results in phosphorylation of PI3K, Akt, and ERK signaling molecules at an early stage of infection, a process mediated by the multistep binding of RVAs outer capsid proteins. Specific inhibitors for PI3K/Akt and MEK/ERK signaling pathways trap the viral particles in late endosome, and acidic replenishment restores and releases them. Moreover, the RVA-induced phosphorylated PI3K, Akt, and ERK directly interact with the subunit E of the V-ATPase proton pump, required for endosomal acidification and RVA uncoating. Understanding how RVA-induced early activation of cellular signaling molecules mediates the V-ATPase-dependent endosomal acidification required for uncoating of viral particles opens up opportunities for targeted interventions against rotavirus entry.
- Subjects :
- 0301 basic medicine
MAPK/ERK pathway
Rotavirus
Viral Diseases
Cell signaling
Physiology
viruses
Signal transduction
ERK signaling cascade
Biochemistry
Viral Packaging
Signaling Molecules
Phosphatidylinositol 3-Kinases
Immune Physiology
Virus Uncoating
Medicine and Health Sciences
Sf9 Cells
Small interfering RNAs
Extracellular Signal-Regulated MAP Kinases
lcsh:QH301-705.5
Late endosome
Cells, Cultured
Immune System Proteins
Chemistry
Signaling cascades
Haplorhini
Hydrogen-Ion Concentration
Cell biology
Precipitation Techniques
Nucleic acids
Infectious Diseases
Phosphorylation
Research Article
lcsh:Immunologic diseases. Allergy
Vacuolar Proton-Translocating ATPases
Signal Inhibition
Immunology
Endosomes
Research and Analysis Methods
Microbiology
Antibodies
Rotavirus Infections
03 medical and health sciences
Virology
Genetics
Immunoprecipitation
Animals
Humans
Non-coding RNA
Molecular Biology
Protein kinase B
PI3K/AKT/mTOR pathway
Rotavirus Infection
Biology and life sciences
Proteins
Viral Replication
Gene regulation
Enzyme Activation
030104 developmental biology
lcsh:Biology (General)
RNA
Parasitology
Capsid Proteins
Cattle
Gene expression
Caco-2 Cells
lcsh:RC581-607
Acids
Proto-Oncogene Proteins c-akt
Subjects
Details
- Language :
- English
- ISSN :
- 15537374 and 15537366
- Volume :
- 14
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- PLoS Pathogens
- Accession number :
- edsair.doi.dedup.....cceb39f2cb23ee906c0a5e0fccf5a563