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Significance Human cytomegalovirus (HCMV) pathogenesis relies upon viral infection of monocytes, as this is the key cell type mediating the hematogenous dissemination of HCMV throughout the host. Here we report that HCMV glycoprotein B surprisingly remains bound to the epidermal growth factor receptor (EGFR) throughout the duration of viral postentry events to induce chronic signaling through EGFR. This chronic EGFR signaling is required to facilitate viral localization to correct trafficking vesicles, as well as promoting nuclear translocation of viral DNA into the monocyte nucleus, both of which are necessary for infection of monocytes. Our data highlight how HCMV regulates the EGFR kinase externally and, uniquely, internally to promote separate and distinct events required for productive infection, spread, and persistence.

HCMV-induced signaling through gB–EGFR engagement is required for viral trafficking and nuclear translocation in primary human monocytes