Identification of picornavirus proteins that inhibit de novo nucleotide synthesis during infection

The picornavirus family includes well-known pathogens for humans and animals, such as enteroviruses (e.g. poliovirus, coxsackievirus, rhinovirus) and cardioviruses (e.g. encephalomyocarditis virus [EMCV] and Saffold virus). Picornaviruses modulate cellular metabolism likely to generate sufficient building blocks for virus replication. Previously, we showed that coxsackievirus B3 (CVB3) and EMCV remodel nucleotide metabolism during infection. Here, we investigated whether this modulation is attributable to specific viral proteins. For this, we studied the modulation of metabolism by several recombinant CVB3 and EMCV viruses in HeLa cells. Using isotope tracing metabolomics with three distinct labels, 13 C 6 -glucose or 13 C 5 / 15 N 2 -glutamine, we reveal that the 2A protease of CVB3 and the Leader protein of EMCV inhibit de novo nucleotide synthesis. Furthermore, we show that nucleotide metabolism is also reprogrammed by CVB3 and EMCV in human induced pluripotent stem cell-derived cardiomyocytes. Our insights are important to increase understanding of picornavirus-host interactions and may lead to novel therapeutic strategies.