CVB3 infection increases purine and pyrimidine metabolites through nucleic acid degradation and nucleotide salvage.

¹³C-glucose isotope tracing study in mock- and CVB3-infected HeLa R19 cells (MOI 5, three replicates; one of three representative experiments is shown). Cells were infected, lysed at 2, 4, 6 or 8 hpi and measured by LC-MS to identify metabolites and quantify the different isotopologues. A) Schematic representation of nucleotide metabolism. Nucleotides can be synthesized de novo, released during degradation of nucleic acids, or recycled (i.e. called the salvage pathway). PRPP, phosphoribosyl pyrophosphate; R5P, ribose-5-phosphate; R1P, ribose-1-phosphate. B) Schematic representation of nucleotide metabolism and nucleotide labeling in ¹³C-glucose tracing studies. Orange = phosphate group; Blue = pentose sugar; Green = nucleobase. C) Isotope distribution of AMP, ADP, ATP, UMP, UDP and UTP in mock and CVB3-infected cells. D) Isotope distribution of dihydroorotate and N-carbamoylaspartate in mock and CVB3-infected cells. The p-values of C) and D) were calculated using linear mixed effect models with an interaction of time and treatment and a random effect of replicate. Afterwards, a contrast analysis was done to calculate the p-values between specific groups. For this analysis, the fractions of all labels were added together and tested whether the total amount of labeling differed between mock and CVB3 infection. *p < 0.05, **p < 0.01, ***p < 0.001.