Abstract:
:Murine gammaherpesvirus 68 (MHV68) is used as a model to study gammaherpesvirus pathogenesis both in tissue culture systems and in vivo. We used a gene-trapping approach to get insight into cellular factors involved in MHV68 infection. By generating a library of gene-trapped CHO cells, we were able to isolate several clones that exhibited various degrees of resistance to MHV68-induced cytopathic effect. Clones that showed the highest degree of resistance were affected at the early stage of the viral cycle, with the vast majority of these clones being deficient for heparan sulfate (HS) expression at the cell surface. Heparan sulfate expression could be restored in all the HS-deficient clones by expression of EXT1, an enzyme that is essential for the biosynthesis of HS. Consistent with the role of HS in viral entry, HS-deficient CHO cells did not support viral internalization. Cell surface heparan sulfate proteoglycans (HSPG) are mostly composed of HS chains attached to two families of core proteins, the transmembrane syndecans and the GPI-anchored glypicans. Treatment of CHO cells with phosphatidylinositol-specific phospholipase C (PI-PLC) did not significantly affect the level of HS expression, indicating that the glypicans are not a major source of HSPG in CHO cells. By contrast, treatment of CHO cells with PMA, a drug known to accelerate syndecan shedding, resulted in a decrease in both HS expression and susceptibility to MHV68; these effects were abolished by TIMP-3, a specific inhibitor of syndecan shedding. All together, our results confirm the essential role of HS in MHV68 infection and identify the syndecans as a major source of HSPG used by the virus as coreceptors to infect CHO cells.
journal_name
Virologyjournal_title
Virologyauthors
Jarousse N,Coscoy Ldoi
10.1016/j.virol.2007.12.004subject
Has Abstractpub_date
2008-04-10 00:00:00pages
376-86issue
2eissn
0042-6822issn
1096-0341pii
S0042-6822(07)00811-2journal_volume
373pub_type
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