African swine fever (ASF) is a highly contagious and lethal disease caused by the African swine fever virus (ASFV). Currently, effective vaccines are not available for the prevention and control of ASF. ASFV is susceptible to mutations as it has a large genome and encodes numerous proteins. In addition to evading the host immune response, ASFV utilizes host proteins to regulate its replication. The ASFV p30 protein is involved in virus internalization into the host cell and is expressed throughout the viral replication cycle, influencing viral replication. This study identified the host proteins that interact with p30 using mass spectrometry analysis. Immunoprecipitation analysis confirmed that the ASFV p30 protein interacted with the host proteins CCAR2 and MATR3, co-localizing with them in the cytoplasm. CCAR2 and MATR3 promoted ASFV replication. Conversely, ASFV infection upregulated the expression of CCAR2 and MATR3 in the host. Thus, the ASFV p30 protein regulates ASFV replication by interacting with CCAR2 and MATR3.