The effect of herpesvirus infection and 2-deoxy- d-glucose on glycosphingolipids in BHK-21 cells

Abstract

The effect of Herpes simplex virus (HSV) infection and 2-deoxy- d-glucose (dGlc) on the content of radiolabeled glycosphingolipids was studied in BHK-21 cells, after a 16 hr pulse with D-[3- 3H]serine and [1- 14C]palmitate. HSV infection caused a stimulated incorporation of the labeled precursors into total glycosphingolipids. The content of newly synthesized hematosides was reduced by 50% and newly synthesized ceramides, ceramide monohexosides, and ceramide dihexosides was increased two-to-threefold, suggesting a simplification of the glycolipid pattern. Treatment of infected and uninfected cells with 10 m M dGLc decreased radioactivity in all glycosphingolipids, whereas the ratio of cerarnide to its glycosylated derivatives increased. Glycosylation of endogenous ceramide was studied in vitro using microsomal fractions and UDP-[ 3H]glucose as the sugar donor. UDP-glucose:ceramide-glucosyltransferase activity was higher in microsomal fractions from HSV-infected cells than in preparations from controls, and treated cells had higher activities than untreated controls. Infection and dGlc treatment increased the number or accessibility of glucose acceptor sites. Thus, dGlc inhibits glycolipid synthesis in control and HSV-infected cells by blocking glycosylation of ceramide; the inhibition of glycolipid synthesis may have a role in virion assembly.