Oncolytic virus hijacks GOT1 and pyrimidinosomes to fuel pyrimidine synthesis for replication in tumor cells.

Oncolytic viruses selectively infect and kill tumor cells, but the metabolic adaptations that support their replication remain incompletely understood. Here, using oncolytic Newcastle disease virus (NDV) as a model, we identify glutamic-oxaloacetic transaminase 1 (GOT1) as a key metabolic enzyme required for efficient viral replication through its dual role in de novo pyrimidine synthesis. In NDV-infected tumor cells, GOT1 promotes aspartate production through the malate-aspartate shuttle to support pyrimidine biosynthesis, while also maintaining NAD(+)/NADH homeostasis to activate the mTOR-S6K-CAD signaling axis and further enhance pyrimidine synthesis. These GOT1-dependent metabolic and signaling adaptations sustain pyrimidine biosynthesis and viral replication. In addition, NDV infection promotes pyrimidinosome assembly, and GOT1 functions as a pyrimidinosome-associated component. Together, these findings reveal a mechanism by which oncolytic NDV rewires host pyrimidine metabolism to support its replication and provide a rationale for metabolic modulation of oncolytic virotherapy.