Ceramide synthase 2 deletion decreases the infectivity of HIV-1

The lipid composition of human immunodefiency virus 1 (HIV-1) virions is enriched in sphingomyelin, but the roles that sphingomyelin or other sphingolipids might play in the HIV-1 replication pathway have not been elucidated. In human cells, sphingolipid levels are regulated by ceramide synthase (CerS) enzymes that produce ceramides, which can be converted to sphingomyelins, hexosylceramides, and other sphingolipids. In many cell types, CerS2, which catalyzes the synthesis of very long chain ceramides, is the major CerS. We have examined how CerS2 deficiency affects the assembly and infectivity of HIV-1. As expected, we observed that very long chain ceramide, hexosylceramide, and sphingomyelin were reduced in CerS2 knockout cells. CerS2 deficiency did not affect HIV-1 assembly or the incorporation of the HIV-1 envelope (Env) protein into virus particles, but it reduced the infectivites of viruses produced in the CerS2-deficient cells. The reduced viral infection levels were dependent on HIV-1 Env, since HIV-1 particles that were pseudotyped with the Vesicular Stomatitis Virus (VSV) glycoprotein (G) did not exhibit reductions in infectivity. Moreover, cell-cell fusion assays demonstrated that the functional defect of HIV-1 Env in CerS2-deficient cells was independent of other viral proteins. Overall, our results indicate that the altered lipid composition of CerS2-deficient cells specifically inhibit the HIV-1 Env receptor binding and/or fusion processes.