In Human Papilloma Virus (HPV)-infected cells p53 protein stability is under the regulation of a complex of HPV-E6 and the cellular E3 ligase E6-Associated Protein (E6AP) which is recruited to promote p53 for proteasomal degradation. This is a key step in HPV-related cancers1. The role of E6AP in non-HPV infected cells is only partially understood. We have previously demonstrated a key role for E6AP in the regulation of a p53 regulator, PML, thereby linked it to the regulation of cell proliferation, cellular stress response2 and cellular senescence3. Herein we investigate the role of E6AP in cell motility invasiveness and metastasis. We found that deficiency in E6AP promotes direct cell migration in-vitro and wound closure in-vivo. Likewise, E6AP deficiency promotes cell invasion in-vitro. Remarkably, our pilot study showed that reconstitution of E6AP expression in triple negative breast cancer cells, which express low levels of E6AP, impairs their invasive capacity in-vitro, represses lung metastatic colonization and constrains metastasis in xenografted tumours. Further, we found that loss or downregulation of E6AP expression in human and mouse cells promotes elongated morphology with enhanced membrane protrusion, filopodia, and actin spikes formation. A downregulation of E6AP in tumour and primary cells regulates also stress fibres formation and focal adhesion turnover. In search for the underlying mechanism we found that in E6AP deficient cells Rac1 is activated, co localises with E6AP, and binds E6AP through its HECT–domain. Furthermore, we found that Rac1 is accumulated and stabilised in cells deficient for E6AP. Our results demonstrate for the first time a key role for E6AP in restraining cancer cell motility and metastasis via the regulation of Rho GTPase signalling.