13C-glucose tracing analysis of IFN-treated A549 cells expressing wild-type SARS-CoV-2 Nsp3-4 or a deISGylation-defective Nsp3-4 mutant protein

ABSTRACT:

Interferon (IFN)-stimulated gene 15 (ISG15) regulates diverse cellular processes, including antiviral immunity, through its conjugation to target proteins (ISGylation). Increasing evidence suggests that ISGylation can also reshape cellular metabolism; however, how viruses counteract ISGylation-mediated metabolic rewiring remains poorly understood. This analysis investigated the impact of the deISGylation activity of the SARS-CoV-2 papain-like protease, which is part of the Nsp3 protein, on type I IFN-driven modulation of cellular glucose metabolism. A549 cells expressing empty vector, wild-type (WT) Nsp3-4 from SARS-CoV-2, or a mutant Nsp3-4 protein that is deficient in PLpro deISGylation activity, were treated with IFNα (or mock-treated) and then subjected to 13C-glucose tracing analysis. Following isotope labeling, intracellular metabolites were extracted and analyzed by LC-MS to assess 13C incorporation into intermediates of glycolysis and the pentose phosphate pathway (PPP). Comparative analysis of the results revealed distinct labeling patterns in glycolytic and PPP metabolites under IFN-stimulated conditions, supporting a role for PLpro deISGylation activity in modulating cellular glucose metabolism. These data provide insight into how SARS-CoV-2 Nsp3 counteracts IFN-induced metabolic rewiring through its deISGylation activity.

INSTRUMENT(S): Liquid Chromatography MS - alternating - hilic

PROVIDER: MTBLS14030 | MetaboLights | 2026-05-05

REPOSITORIES: MetaboLights