Multilevel proteomics reveals common and specific host perturbations by Monkeypox and Vaccinia viruses

The global spread of Monkeypox virus (MPXV) has resulted in the urgent need for an in-depth molecular characterization of the virus infection. Multiple omics studies on Vaccinia virus have extended our knowledge of poxvirus pathophysiology (REFs). Nevertheless, there are no comparative studies that would include an in-depth comparison of MPXV with other poxviruses in the context of molecular biology. Here, we report a comparative time-resolved proteomic study of MPXV and vaccinia viruses and a concurrent multi-omics study of MPXV infection in primary human cells. Using state-of-the-art proteomics, we profiled the virus-host interplay on the transcriptome, proteome and phosphoproteome levels in primary human foreskin fibroblasts. Pathway analysis in combination with projecting the gathered data onto the global network of host-signaling interactions revealed crosstalk between the perturbations at different levels, enabling identification of distinct and common molecular mechanisms of poxviruses. The NF-κB pathway, a signaling pathway governing immunomodulation and inflammation, was unexpectedly activated by MPXV but not VacV, while MPXV exhibited an astounding resistance to the effects of antiviral interferon response, potentially explaining the unique pathogenicity hallmarks of the monkeypox.