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J. Virol., 06 1995, 3290-3298, Vol 69, No. 6
BW Banfield, Y Leduc, L Esford, K Schubert and F Tufaro
A novel mouse L-cell mutant cell line defective in the biosynthesis of
glycosaminoglycans was isolated by selection for cells resistant to herpes
simplex virus (HSV) infection. These cells, termed sog9, were derived from
mutant parental gro2C cells, which are themselves defective in heparan
sulfate biosynthesis and 90% resistant to HSV type 1 (HSV-1) infection
compared with control L cells (S. Gruenheid, L. Gatzke, H. Meadows, and F.
Tufaro, J. Virol. 67:93-100, 1993). In this report, we show that sog9 cells
exhibit a 3-order-of-magnitude reduction in susceptibility to HSV-1
compared with control L cells. In steady-state labeling experiments, sog9
cells accumulated almost no [35S]sulfate-labeled or
[6-3H]glucosamine-labeled glycosaminoglycans, suggesting that the
initiation of glycosaminoglycan assembly was specifically reduced in these
cells. Despite these defects, sog9 cells were fully susceptible to
vesicular stomatitis virus (VSV) and permissive for both VSV and HSV
replication, assembly, and egress. HSV plaques formed in the sog9
monolayers in proportion to the amount of input virus, suggesting the block
to infection was in the virus entry pathway. More importantly, HSV-1
infection of sog9 cells was not significantly reduced by soluble heparan
sulfate, indicating that infection was glycosaminoglycan independent.
Infection was inhibited by soluble gD-1, however, which suggests that
glycoprotein gD plays a role in the infection of this cell line. The block
to sog9 cell infection by HSV-1 could be eliminated by adding soluble
dextran sulfate to the inoculum, which may act by stabilizing the virus at
the sog9 cell surface. Thus, sog9 cells provide direct genetic evidence for
a proteoglycan-independent entry pathway for HSV-1, and results with these
cells suggest that HSV-1 is a useful reagent for the direct selection of
novel animal cell mutants defective in the synthesis of cell surface
proteoglycans.
Copyright © 1995, American Society for Microbiology
Sequential isolation of proteoglycan synthesis mutants by using herpes simplex virus as a selective agent: evidence for a proteoglycan- independent virus entry pathway
Department of Microbiology and Immunology, University of British Columbia, Vancouver, Canada.
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