Leptospira interrogans is the causative agent of leptospirosis, a potentially severe and life-threatening zoonotic infectious disease. One of the symptoms associated with severe leptospirosis is acute kidney failure, with detachment of renal tubule epithelial cells (RPTECs) from the basement membrane, which can develop into chronic kidney disease. The kidney intrinsically possesses the ability to self-repair after acute injury through collective cell migration, which involves the movement of cells as a cohesive unit. L. interrogans induces the disassembly of adherens junctions, which can be prevented by inhibitors of the eukaryotic proteolytic pathways. However, the impact on epithelial repair processes have not been investigated yet. In this study, we performed a proteome-wide analysis of protein ubiquitination in infected-RPTECs and identified ARL2, a protein involved in microtubule dynamics. Immunofluorescence analysis showed that L. interrogans induces a proteasomal-dependent disruption of the microtubule/F-actin cytoskeletal network. Consequently, an aberrant collective cell migration was observed in a scratch assay, which was improved by proteasomal inhibition. Our data suggested that L. interrogans alters a circuit integrating mechanosensors at cell–cell contacts and the supracellular F-actin/microtubule cytoskeleton network. Proteasome inhibition prevents the disruption of this circuit and might improve RPTECs' repair responses. Our findings emphasize the role of the Ubiquitinin Proteasome System in leptospiral pathogenesis as the basis for novel treatment strategies to prevent renal damage.