Coronavirus Nucleocapsid Proteins Hijack Host Protein Kinase A Catalytic Subunit α into Nucleus to Evade from STAT1 Signaling

Abstract The role of the cAMP/PKA pathway in antiviral innate immunity, including during coronavirus infections, remains unclear. We discovered coronavirus N proteins initiate cAMP-ADCY10-PKA cascade. Cytoplasmic PKA activates STAT1 independently of canonical JAK/TYR2 signaling. Coronavirus N proteins, via a conserved arginine (e.g., PEDV R58, SARS-CoV-2 R92), directly bind and sequester PKA Cα into the nucleus to evade STAT1 activation. Using PEDV as a model, mutant viruses with N mutNLS and N R58A were generated. Wildtype PEDV infection suppressed STAT1 activation in cells expressing PKA Cα/PKA Cα S339A deficient in STAT1 phosphorylation. However, in rXS0101 mutNLS - or rXS0101 R58A -infected cells, only PKA Cα S339A expression inhibited STAT1 activation. Downregulation of STAT1 activation is accompanied with increased viral replication. This study first elaborates that PKA Cα activates STAT1 in the cytoplasm of infected cells distinctly from canonical JAK/STAT1 signaling, while coronaviruses evade this antiviral response by sequestering PKA Cα into the nucleus via direct N protein interaction.