A receptor switch between discoidin domain receptors underlies orthobunyavirus entry and host range

Orthobunyaviruses, an important group of arboviruses with broad vector and host ranges, pose a persistent global public health threat, yet their cellular entry and cross-species transmission remain poorly understood. Ebinur Lake virus (EBIV), a recently identified orthobunyavirus detected in multiple countries and hosts, demonstrates potential zoonotic and spillover relevance. Using a genome-wide CRISPR screen, we identify discoidin domain receptor 2 (DDR2) as an entry receptor for EBIV. Knockout and ectopic expression confirm that DDR2 promotes EBIV infectivity by facilitating viral binding and internalization. The discoidin (DS) domain of DDR2 directly interacts with EBIV particles at the Gc stalk region. A soluble DS decoy or anti-DDR2 DS antibody potently inhibits EBIV infection in cell lines and primary neural cells. The DS decoy or Ddr2 knockout protects mice from lethal EBIV challenge, significantly reducing viral loads. DDR2 homologs from diverse mammals and birds bind EBIV Gc and support infection, underscoring a broad host range. Notably, we demonstrate a receptor switch among phylogenetically related orthobunyaviruses: human-infecting Germiston virus (GERV) also utilizes DDR2, while Bozo virus (BOZOV) employs DDR1. These findings establish discoidin domain receptors as functional entry receptors for orthobunyaviruses, revealing a mechanism for broad host tropism and providing insights for antiviral strategies. In this study, the authors identify DDR2 as an entry receptor for Ebinur Lake virus, show that it promotes infection in vitro and in vivo, and reveal a receptor switch among related orthobunyaviruses that underlies broad host range and zoonotic potential.