Global Landscape of Human Kinase Motifs in Viral Proteomes

The successful establishment of infection relies on an ability to sense and adapt to the host signaling state. One key mechanism of virus-host sensing is host-mediated post-translational modifications of viral proteins. While viral protein phosphorylation by host kinases is known to modulate viral functions, the global prevalence of kinase motifs across diverse viruses, and the signaling pathways they reflect, remains to be systematically explored. Here, we annotated human kinase motifs in 1,505 viral proteomes and uncovered enriched motifs in viral proteins that diverged from patterns observed in human proteins. Integration of our findings with 21,606 viral protein structures and deep mass spectrometry phosphoproteomics of infected human cells revealed that surface-accessible residues were preferentially phosphorylated and exhibited greater kinase specificity compared to buried sites. Virus-enriched motifs mapped predominantly to stress, inflammation, and cell cycle pathways—key signaling hubs dysregulated during infection that are central to the virus-host arms race—most strikingly for Flaviviridae , Togaviridae , Herpesviridae, and Retroviridae families. Temporal phosphoproteomic profiling of host kinase activity during alphavirus infection revealed dynamic patterns of stress kinase activation and viral protein phosphorylation, and the inhibition of MAP kinases reduced viral replication and phosphorylation at viral motifs with specificity for ERK and JNK kinases. Our findings suggest that viruses have evolved as biosensors of the host signaling state to optimize their life cycles, revealing new antiviral opportunities aimed at disrupting virus decision-making by manipulating host signaling cues.