Delta-like 1-Lysine613 regulates notch signaling

► Seventeen conserved lysine residues in the cytoplasmic tail of Delta like1 (Dll1) were identified and mutated. ► Notch signaling, ubiquitination, endocytic recycling, lipid raft localization and Notch binding functions were examined. ► K613 was identified as important for Notch signaling. ► K613R altered the multiubiquitination profile of Dll1. ► K613R did not alter endocytosis or stability of Dll1 but did alter binding to Notch1 and lipid raft localization.