Dysregulation of autophagy in murine fibroblasts resistant to HSV-1 infection.
The mouse L cell mutant, gro29, was selected for its ability to survive infection by herpes simplex virus type 1 (HSV-1). gro29 cells are fully susceptible to HSV-1 infection, however, they produce 2000-fold less infectious virus than parental L cells despite their capacity to synthesize late viral gene products and assemble virions. Because productive HSV-1 infection is presumed to result in the death of the host cell, we questioned how gro29 cells might survive infection. Using time-lapse video microscopy, we demonstrated that a fraction of infected gro29 cells survived infection and divided. Electron microscopy of infected gro29 cells, revealed large membranous vesicles that contained virions as well as cytoplasmic constituents. These structures were reminiscent of autophagosomes. Autophagy is an ancient cellular process that, under nutrient deprivation conditions, results in the degradation and catabolism of cytoplasmic components and organelles. We hypothesized that enhanced autophagy, and resultant degradation of virions, might explain the ability of gro29 to survive HSV-1 infection. Here we demonstrate that gro29 cells have enhanced basal autophagy as compared to parental L cells. Moreover, treatment of gro29 cells with 3-methyladenine, an inhibitor of autophagy, failed to prevent the formation of autophagosome-like organelles in gro29 cells indicating that autophagy was dysregulated in these cells. Additionally, we observed robust co-localization of the virion structural component, VP26, with the autophagosomal marker, GFP-LC3, in infected gro29 cells that was not seen in infected parental L cells. Collectively, these data support a model whereby gro29 cells prevent the release of infectious virus by directing intracellular virions to an autophagosome-like compartment. Importantly, induction of autophagy in parental L cells did not prevent HSV-1 production, indicating that the relationship between autophagy, virus replication, and survival of HSV-1 infection by gro29 cells is complex.
Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario, Canada.
Valerie
V
Le Sage
Bruce W
BW
Banfield
D000818:Animals; D001343:Autophagy; D036022:Capsid Proteins / Q000378:metabolism; D002470:Cell Survival; D002522:Chlorocebus aethiops; D022162:Cytoplasmic Vesicles / Q000821:virology; D004268:DNA-Binding Proteins / Q000378:metabolism; D005347:Fibroblasts / Q000276:immunology / Q000821:virology*; D018259:Herpesvirus 1, Human / Q000378:metabolism*; D007739:L Cells; D051379:Mice; D010641:Phenotype; D010766:Phosphorylation; D021381:Protein Transport; D014157:Transcription Factors / Q000378:metabolism; D014709:Vero Cells; D014771:Virion / Q000648:ultrastructure; D014779:Virus Replication
D036022:Capsid Proteins; D004268:DNA-Binding Proteins; C103652:Elf2 protein, mouse; D014157:Transcription Factors; C077409:capsid protein VP26, herpes simplex virus type 1