MUC1 is normally expressed on the apical surface of epithelial cells, where its highly extended mucin-like structure serves a protective role by modulating clearance or retention of secreted mucins and by providing a scaffold for the presentation of glycans that are recognized by bacteria and viruses (1-9). In tumors of epithelial origin, cell polarity is lost, and MUC1 expression on all cell surfaces contributes to an aggressive tumor phenotype; the extended peptide core inhibits cell-cell and cell-matrix interactions, whereas the presence of specific glycan structures such as sialyl-Le X and sialyl-Le a can act as ligands for selectin-like molecules on endothelial cells and thereby enhance metastasis (10 -13).The anti-adhesive property of MUC1 is also enhanced by its ability to compete with the cell adhesion molecule E-cadherin for binding of cytoplasmic -catenin, an important link in the maintenance of actin interactions with the adherins junctions of epithelia (14, 15). In fact, loss of E-cadherin and aberrant localization of both -catenin and MUC1 correlate with an aggressive tumor phenotype and a poor prognosis for the patient (16, 17). The binding of -catenin to MUC1 is regulated by phosphorylation at adjacent sites by glycogen synthase kinase-3, Src family kinases, the epidermal growth factor receptor, or protein kinase C␦ (15, 18 -22). MUC1 is autocatalytically cleaved within the SEA (sea urchin sperm protein, enterokinase, and agrin) domain in the endoplasmic reticulum, but the larger mucin-like subunit remains tightly associated with the small transmembrane subunit (23-25). Using antibodies directed against a peptide corresponding to the MUC1 small subunit C terminus, researchers have reported that (i) treatment of human ZR-75-1 breast cancer cells with the ErbB ligand heregulin targets a complex of ␥-catenin and MUC1 to the nucleus; (ii) overexpression of MUC1 in pancreatic cancer cell lines targets -catenin and MUC1 to the nucleus; (iii) activation of Lyn kinase in multiple myeloma cells with interleukin-7 targets a complex of -catenin and MUC1 to the nucleus; and (iv) MUC1 is targeted to mitochondria when HCT116 colon carcinoma cells overexpressing MUC1 are stimulated with heregulin (20, 26 -28). Although nuclear and cytoplasmic -catenins were observed by one research group in breast cancer patients (17), others found -catenin and MUC1 only in the cytoplasm and plasma membrane in both human breast cancer samples and a spontaneous mouse model of breast cancer (29 -31).The mechanism for nuclear or mitochondrial targeting of the MUC1 small subunit is unknown, but delivery of the subunit to any intracellular compartment is likely dependent on its endocytosis from the cell surface. We have reported previously that MUC1 is internalized faster with shorter glycans (32), a feature of MUC1 expressed in several human breast tumor cell lines (33-36). Replacement of the extended ectodomain of MUC1 with that of Tac (interleukin-2 receptor ␣-subunit) also enhances endocytosis; and using site-specific mut...