Blockade of pan-viral propagation by inhibition of host cell PNPT1.

• Viruses exploit a common survival strategy of inducing host cell polyribonucleotide nucleotidyltransferase 1 (PNPT1). • PNPT1 mitigates the integrated stress response mediated by mitochondrial double-stranded RNAs (mt-dsRNAs) during viral propagation. • PNPT1 depletion inhibits pan-viral propagation by activating the mt-dsRNA-RNA-activated protein kinase-eukaryotic initiation factor 2α signal axis. • The PNPT1 inhibitor, lanatoside C, was identified as a potent blocker of pan-viral propagation. For successful viral propagation within infected cells, the virus needs to overcome the cellular integrated stress response (ISR), triggered during viral infection, which, in turn, inhibits general protein translation. This paper reports a tactic employed by viruses to suppress the ISR by upregulating host cell polyribonucleotide nucleotidyltransferase 1 (PNPT1). The propagation of adenovirus, murine cytomegalovirus and hepatovirus within their respective host cells induces PNPT1 expression. Notably, when PNPT1 is knocked down, the propagation of all three viruses is prevented. Mechanistically, the inhibition of PNPT1 facilitates the relocation of mitochondrial double-stranded RNAs (mt-dsRNAs) to the cytoplasm, where they activate RNA-activated protein kinase (PKR). This activation leads to eukaryotic initiation factor 2α (eIF2α) phosphorylation, resulting in the suppression of translation. Furthermore, by scrutinizing the PNPT1 recognition element and screening 17,728 drugs and bioactive compounds approved by the US Food and Drug Administration, lanatoside C was identified as a potent PNPT1 inhibitor. This compound impedes the propagation of adenovirus, murine cytomegalovirus and hepatovirus, and suppresses production of the severe acute respiratory syndrome coronavirus-2 spike protein. These discoveries shed light on a novel strategy to impede pan-viral propagation by activating the host cell mt-dsRNA-PKR-eIF2α signalling axis. [ABSTRACT FROM AUTHOR]