Viruses are obligatory intracellular microbes that exploit the host machinery to meet their biosynthetic demands. Targeting a universal host protein exploited by most viruses would be a game-changing strategy that offers broad-spectrum solution and rapid pandemic control. Along this direction, (i) we found a common YxxØ-motif of multiple viruses that exploits host AP2M1 for intracellular trafficking. A library chemical compound, ACA, was identified to interrupt AP2M1-virus interaction and exhibit potent antiviral efficacy against a number of viruses in vitro and in vivo; (ii) utilizing influenza A virus (IAV) and SARS-CoV-2 as models, we demonstrated that both viral nucleoprotein (NP) harnessed host transcriptional corepressor TLE1 for enhancing viral polymerase activity while suppressing cellular innate response. NP-TLE1 recognition was realized via a conserved bHLH-like motif. Blockage of NP-TLE1 interaction by a peptide disruptor AH49A provides antiviral protection in vitro and in vivo. Our studies reveal evolutionarily conserved mechanism of host-virus interactions with broad relevance to human viral infections, which represents potential strategies for the development of antiviral agents.