Modulation of Zika virus replication via glycosphingolipids

The enveloped positive-sense RNA viruses including Zika virus (ZIKV) need host lipids to successfully replicate. The nature of the lipids and the replication step(s) where lipids are utilized often vary amongst viruses. In this study, we demonstrate that ZIKV particle envelope is significantly enriched in distinct sphingolipid species. To determine the role of sphingolipids in ZIKV replication, we leveraged a panel of sphingolipid-deficient cell lines. Notably, knockout of glucosylceramide and lactosylceramide synthase encoding genes (GCSKO; B4G5KO) resulted in a marked decrease in ZIKV titers. GCSKO or pharmacological inhibition of GCS also led to a significant decrease in ZIKV genome replication. Further analysis indicated that GCSKO reduced intracellular virus titers but had minimal impact on ZIKV binding. Restoration of B4G5 expression in B4G5KO cells or supplementing PDMP-treated cells with glucosylceramide led to a significant rescue of ZIKV replication. Altogether, our findings suggest that ZIKV needs glycosphingolipids to facilitate virus replication. •Quantitative analysis of partially purified ZIKV particles showed an enrichment of sphingolipid and glycosphingolipid species.•Transcription activator-like effector nucleases (TALENs)-based knockout of glucosylceramide synthase (GCS) gene and pharmacological inhibition of GCS activity resulted in a significant reduction in ZIKV titers and genome replication.•Supplementing cells with glucosylceramide glycosphingolipid led to a rescue of ZIKV replication in cells treated with a pharmacological inhibitor of GCS activity.•SPharmacological inhibition of GCS activity at both early and late stages of ZIKV infection cycle led to a significant reduction in ZIKV titers, implying a role for glycosphingolipids in ZIKV genome replication and post replication steps.•Our findings reveal the importance of GCS, and its related glycosphingolipids, in ZIKV genome replication and post replication steps.