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Deletion of the Pseudorabies Virus gE/gI-US9p complex disrupts kinesin KIF1A and KIF5C recruitment during egress, and alters the properties of microtubule-dependent transport in vitro
- Source :
- PLoS Pathogens, Vol 16, Iss 6, p e1008597 (2020), PLoS Pathogens
- Publication Year :
- 2020
- Publisher :
- Public Library of Science (PLoS), 2020.
-
Abstract
- During infection of neurons by alphaherpesviruses including Pseudorabies virus (PRV) and Herpes simplex virus type 1 (HSV-1) viral nucleocapsids assemble in the cell nucleus, become enveloped in the cell body then traffic into and down axons to nerve termini for spread to adjacent epithelia. The viral membrane protein US9p and the membrane glycoprotein heterodimer gE/gI play critical roles in anterograde spread of both HSV-1 and PRV, and several models exist to explain their function. Biochemical studies suggest that PRV US9p associates with the kinesin-3 motor KIF1A in a gE/gI-stimulated manner, and the gE/gI-US9p complex has been proposed to recruit KIF1A to PRV for microtubule-mediated anterograde trafficking into or along the axon. However, as loss of gE/gI-US9p essentially abolishes delivery of alphaherpesviruses to the axon it is difficult to determine the microtubule-dependent trafficking properties and motor-composition of Δ(gE/gI−US9p) particles. Alternatively, studies in HSV-1 have suggested that gE/gI and US9p are required for the appearance of virions in the axon because they act upstream, to help assemble enveloped virions in the cell body. We prepared Δ(gE/gI-US9p) mutant, and control parental PRV particles from differentiated cultured neuronal or porcine kidney epithelial cells and quantitated the efficiency of virion assembly, the properties of microtubule-dependent transport and the ability of viral particles to recruit kinesin motors. We find that loss of gE/gI-US9p has no significant effect upon PRV particle assembly but leads to greatly diminished plus end-directed traffic, and enhanced minus end-directed and bidirectional movement along microtubules. PRV particles prepared from infected differentiated mouse CAD neurons were found to be associated with either kinesin KIF1A or kinesin KIF5C, but not both. Loss of gE/gI-US9p resulted in failure to recruit KIF1A and KF5C, but did not affect dynein binding. Unexpectedly, while KIF5C was expressed in undifferentiated and differentiated CAD neurons it was only found associated with PRV particles prepared from differentiated cells.<br />Author summary Alphaherpesviruses including Pseudorabies virus (PRV) and Herpes simplex virus type 1 (HSV-1) are pathogens of the nervous system. They replicate in the nerve cell body then travel great distances along axons to reach nerve termini and spread to adjacent epithelial cells. Virally encoded membrane proteins US9p and the heterodimer gE/gI play critical roles in anterograde spread for both HSV-1 and PRV, however there is disagreement over their roles. Here we compared mutant PRV virions lacking gE/gI-US9p with a parental control strain, and quantitated the efficiency of virion assembly, the properties of microtubule-dependent transport and the ability of viral particles to recruit kinesins. We find that loss of gE/gI-US9p has no effect upon PRV particle assembly but that mutant virions demonstrate greatly reduced transport to the plus-ends of microtubules, enhanced minus-end traffic and greater frequency of reversal in their motion. These changes are accompanied by loss of both kinesin KIF1A and kinesin KIF5C from the mutant, motors that we find associated with distinct populations of the parental virus. We propose a model in which gE/gI-US9p binds KIF1A to ensure efficient plus end-mediated transport and delivery of PRV particles to a location at which KIF5C is subsequently recruited.
- Subjects :
- Cellular differentiation
viruses
Kinesins
Microtubules
Axonal Transport
Biochemistry
Virions
Nerve Fibers
Viral Envelope Proteins
Animal Cells
Axon
Biology (General)
Virus Release
Cytoskeleton
Neurons
0303 health sciences
biology
Chemistry
030302 biochemistry & molecular biology
Intracellular Signaling Peptides and Proteins
Microtubule Motors
Cell Differentiation
Herpesvirus 1, Suid
Cell biology
medicine.anatomical_structure
Cell Processes
Virion assembly
Kinesin
Cellular Structures and Organelles
Cellular Types
Neuronal Differentiation
Research Article
Imaging Techniques
QH301-705.5
Lipoproteins
Immunology
Biological Transport, Active
Viral Structure
Research and Analysis Methods
Microbiology
Cell Line
Viral Proteins
03 medical and health sciences
Molecular Motors
Microtubule
Virology
Fluorescence Imaging
Genetics
medicine
Animals
Humans
Molecular Biology
030304 developmental biology
Pseudorabies
Biology and Life Sciences
Proteins
Cell Biology
Viral membrane
RC581-607
Axons
Cytoskeletal Proteins
Membrane glycoproteins
Cellular Neuroscience
biology.protein
Axoplasmic transport
Parasitology
Immunologic diseases. Allergy
Gene Deletion
Developmental Biology
Neuroscience
Subjects
Details
- Language :
- English
- ISSN :
- 15537374 and 15537366
- Volume :
- 16
- Issue :
- 6
- Database :
- OpenAIRE
- Journal :
- PLoS Pathogens
- Accession number :
- edsair.doi.dedup.....5fd0a65d1520fefc98e2c78bf40137dc