5-Propyl-2-deoxyuridine induced interference with glycosylation in herpes simplex virus infected cells. Nature of PdU-induced modifications of N-linked glycans.
In herpes simplex virus-infected (HSV) cells, the antiviral nucleoside analogue 5-n-propyl-2'-deoxyuridine (PdU) may, under certain circumstances, induce a pattern of interference with late steps in formation of N-linked glycans, resulting in increased availability of viral glycoproteins for neutralizing antibodies. The PdU-induced changes in N-linked glycans, released by pronase digestion of the HSV-specified glycoprotein gC-1, were investigated by using lectin affinity chromatography and Bio-Gel P6 gel filtration of glycans, radiolabelled with [3H]galactose or [3H]glucosamine. PdU-treatment of HSV-infected cells totally inhibited addition of sialic acid and reduced the amount of galactose incorporated into N-linked glycans by 70%. In addition, the PDU-treatment caused a decrease in oligosaccharides with affinity for Phaseoulus vulgaris leuco-agglutinin and erythro-agglutinin, and an increase in Lens culinaris lectin (LCA)-binding oligosaccharides, suggesting a PdU-induced shift from multi-branched to moderately branched structures. This shift was also found in HSV-infected B16 mouse melanoma cells, where the large content of multi-branched oligosaccharides contributes to the metastatic potential. The LCA-binding glycans from PdU-treated cells were smaller and contained less galactose units than corresponding structures from untreated cells. In a cell-free system, PdU 5'-monophosphate inhibited the translocation of UDP-GlcNAc, and, to a smaller extent, also the translocation of UDP-galactose into Golgi vesicles, suggesting that nucleotide sugar translocation is one important target for the PdU-induced interference with glycosylation in HSV-infected cells.
Department of Clinical Virology, University of Göteborg, Sweden.
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S
Olofsson
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Sjöblom
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K
Hellstrand
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D
Shugar
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Clairmont
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Hirschberg
D000373:Agglutinins / Q000378:metabolism; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D001973:Bromodeoxyuridine / Q000031:analogs & derivatives / Q000494:pharmacology; D002460:Cell Line; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003857:Deoxyuridine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006031:Glycosylation / Q000187:drug effects; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D007223:Infant; D037102:Lectins; D008546:Melanoma, Experimental; D051379:Mice; D009439:Neuraminidase / Q000494:pharmacology; D009702:Nucleoside Diphosphate Sugars / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism*; D018139:Simplexvirus / Q000187:drug effects* / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins / Q000378:metabolism
D000373:Agglutinins; D000998:Antiviral Agents; D037102:Lectins; D009702:Nucleoside Diphosphate Sugars; D011134:Polysaccharides; D014759:Viral Envelope Proteins; C038535:glycoprotein gC, herpes simplex virus type 1; C024068:5-propyl-2'-deoxyuridine; C020235:brivudine; D009439:Neuraminidase; D001973:Bromodeoxyuridine; D003857:Deoxyuridine