The intracellular parasite Toxoplasma gondii enhances its dissemination to distant organs by hijacking 22 infected leukocytes via a Trojan Horse mechanism. Upon infecting dendritic cells (DCs), Toxoplasma 23 induces a hypermigratory phenotype characterized by podosome dissolution and formation of F-actin 24 stress fibers. We previously showed that these cytoskeletal changes depend on the effector protein 25 Toxoplasma WAVE complex-interacting protein (TgWIP) secreted from parasites to infected leukocytes. 26 Here, we identify the host adaptor proteins Non-catalytic region of tyrosine kinase adaptor protein 1 and 2 27 (Nck1/2) and Growth factor receptor-bound protein 2 (Grb2) as direct TgWIP interactors. TgWIP mainly 28 uses two distinct proline-rich regions (PRRs) to interact with Nck1 and Grb2. Mutating these PRRs 29 abrogates TgWIP binding to Nck1 and Grb2 and diminishes podosome dissolution and DC hypermotility. 30 Furthermore, we show that TgWIP directly interacts with the actin nucleation promoting factor WAVE 31 Regulatory Complex (WRC) via a WRC-interacting receptor sequence (WIRS). Disrupting this 32 interaction also influences actin cytoskeletal remodeling and DC hypermotility. Collectively, our data 33 reveal that TgWIP directly interacts with multiple actin regulators, including Nck1, Grb2, and the WRC, 34 to remodel the actin cytoskeleton of the host cells, elucidating a key mechanism that Toxoplasma exploits 35 to enhance host cell migration and dissemination.