African swine fever virus (ASFV) is a large DNA virus causing a highly contagious disease in domestic and wild boar, for which no treatment is available. ASFV vaccine development is hindered by large gaps in knowledge considering virus protein function and virus-host interaction. This study identifies the interaction between two ASFV proteins, MGF360-21R and A151R, and a host barrier-to-autointegration factor 1 (BANF1). We showed that proteins MGF360-21R and A151R can inhibit the induction of the IFN response. Additionally, BANF1 was determined as an important factor in ASFV infection, as the downregulation of BANF1 reduces virus replication. These results uncover a novel function of viral proteins MGF360-21R and A151R and extend our understanding of complex interaction between virus and host, providing insights for developing a vaccine to prevent and control ASFV.