The Chikungunya virus (CHIKV) is a single positive-stranded RNA virus transmitted by mosquitoes. More than 40 countries have reported cases of CHIKV infection to varying degrees. Currently, there is no licensed specific antiviral drug or vaccine for CHIKV in the world. In this study, we have identified host protein interactors of CHIKV nsP4 proteins in 293T cells and the major cellular pathways involved in the interaction between host factors and CHIKV nsp4 using Co-Immunoprecipitation and GST Pull-down followed by high-throughput quantitative mass spectrometry.