The study investigates the mechanisms of clathrin-independent endocytosis of ubiquitinated cargos, focusing on the epidermal growth factor receptor (EGFR). At low doses of EGF, EGFR is internalized primarily through the clathrin pathway and remains unubiquitinated. However, at higher EGF concentrations, a significant fraction of the receptor is internalized via a clathrin-independent, lipid raft-dependent route, becoming ubiquitinated. This non-clathrin internalization depends on the interaction of ubiquitinated EGFR with proteins containing the Ub-interacting motif (UIM), such as eps15, eps15R, and epsin. The study uses a chimera of EGFR fused to a mutant Ub (EGFR/Ubmut) to demonstrate that ubiquitination is sufficient for internalization. Pharmacological and genetic approaches further reveal that eps15/eps15R and epsin are essential for non-clathrin internalization, while clathrin is required for clathrin-dependent internalization at low EGF concentrations. The findings highlight the role of UIM proteins in coupling ubiquitinated cargo to non-clathrin endocytosis and suggest that ubiquitination of endocytic machinery, rather than cargo, is crucial for internalization into coated pits.The study investigates the mechanisms of clathrin-independent endocytosis of ubiquitinated cargos, focusing on the epidermal growth factor receptor (EGFR). At low doses of EGF, EGFR is internalized primarily through the clathrin pathway and remains unubiquitinated. However, at higher EGF concentrations, a significant fraction of the receptor is internalized via a clathrin-independent, lipid raft-dependent route, becoming ubiquitinated. This non-clathrin internalization depends on the interaction of ubiquitinated EGFR with proteins containing the Ub-interacting motif (UIM), such as eps15, eps15R, and epsin. The study uses a chimera of EGFR fused to a mutant Ub (EGFR/Ubmut) to demonstrate that ubiquitination is sufficient for internalization. Pharmacological and genetic approaches further reveal that eps15/eps15R and epsin are essential for non-clathrin internalization, while clathrin is required for clathrin-dependent internalization at low EGF concentrations. The findings highlight the role of UIM proteins in coupling ubiquitinated cargo to non-clathrin endocytosis and suggest that ubiquitination of endocytic machinery, rather than cargo, is crucial for internalization into coated pits.