Targeting cysteine-mediated dimerization of the MUC1-C oncoprotein in             human cancer cells.
The MUC1 heterodimeric protein is aberrantly overexpressed in diverse human carcinomas and contributes to the malignant phenotype. The MUC1-C transmembrane subunit contains a CQC motif in the cytoplasmic domain that has been implicated in the formation of dimers and in its oncogenic function. The present study demonstrates that MUC1-C forms dimers in human breast and lung cancer cells. MUC1-C dimerization was detectable in the cytoplasm and was independent of MUC1-N, the N-terminal mucin subunit that extends outside the cell. We show that the MUC1-C cytoplasmic domain forms dimers in vitro that are disrupted by reducing agents. Moreover, dimerization of the MUC1-C subunit in cancer cells was blocked by reducing agents and increased by oxidative stress, supporting involvement of the CQC motif in forming disulfide bonds. In support of these observations, mutation of the MUC1-C CQC motif to AQA completely blocked MUC1-C dimerization. Importantly, this study was performed with MUC1-C devoid of fluorescent proteins, such as GFP, CFP and YFP. In this regard, we show that GFP, CFP and YFP themselves form dimers that are readily detectable with cross-linking agents. The present results further demonstrate that a cell-penetrating peptide that targets the MUC1-C CQC cysteines blocks MUC1-C dimerization in cancer cells. These findings provide definitive evidence that: i) the MUC1-C cytoplasmic domain cysteines are necessary and sufficient for MUC1-C dimerization, and ii) these CQC motif cysteines represent an Achilles' heel for targeting MUC1-C function.
Genus Oncology, Boston, MA, USA.
Deepak
D
Raina
Rehan
R
Ahmad
Hasan
H
Rajabi
Govind
G
Panchamoorthy
Surender
S
Kharbanda
Donald
D
Kufe
D020816:Amino Acid Motifs; D000970:Antineoplastic Agents / Q000378:metabolism* / Q000494:pharmacology; D003545:Cysteine; D003593:Cytoplasm / Q000378:metabolism; D019281:Dimerization; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D045325:HCT116 Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018396:Mucin-1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism*; D009842:Oligopeptides / Q000378:metabolism* / Q000494:pharmacology; D018384:Oxidative Stress; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D019163:Reducing Agents / Q000494:pharmacology; D014162:Transfection
D000970:Antineoplastic Agents; D004220:Disulfides; D008164:Luminescent Proteins; C060192:MUC1 protein, human; D018396:Mucin-1; D009842:Oligopeptides; D019163:Reducing Agents; D003545:Cysteine