Myxovirus resistance protein A(MxA), one of the dynamin superfamily of large guanosine triphosphatase and a classical interferon stimulated gene (ISG) induced by type I interferons (IFNs), plays antiviral role in various virus infections. However, the effect of MxA on Zika virus (ZIKV) infection and its underlying mechanism remain elusive. In this study, we aimed to explore the role of MxA in ZIKV infection and its potential mechanisms. MxA overexpression was achieved by transfection with plasmid. The levels of MxA expression and ZIKV replication were assayed by both qRT-PCR and western blot. The activation status of Jak/STAT signaling pathway was evaluated at three levels: phosphorylation of STAT1 and STAT2(p-STAT1, p-STAT2) (western blot), activity of interferon sensitive response element (ISRE) (dual luciferase reporter gene assay), and the expression levels of ISGs (qRT-PCR). Our results showed that MxA overexpression inhibited ZIKV replication with no effect on virus entry. The expression levels of retinoic acid inducible gene I (RIG-I), melanoma differentiation-associated gene-5(MDA5), Toll-like receptor3(TLR3) and interferon regulatory Factor 3(IRF3), as well as IFNα and IFNβ, were increased in parallel with MxA upregulation. Interestingly, the inhibitory effect of MxA on ZIKV replication was abolished in type I IFN receptor (IFNAR) deficient cells (U5A). These data collectively supported that MxA inhibits ZIKV replication through activation of the type I IFN signaling pathway.