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J. Biol. Chem., Vol. 275, Issue 44, 34672-34680, November 3, 2000

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Identification of the Elements Regulating the Expression of the Cell Adhesion Molecule MCAM/MUC18
LOSS OF AP-2 IS NOT REQUIRED FOR MCAM EXPRESSION IN MELANOMA CELL LINES^*

C. Susan Mintz-Weber and Judith P. Johnson

From the Institute for Immunology, University of Munich, Munich 80336, Germany

The cell adhesion molecule melonoma cell adhesion molecule (MCAM)/MUC18/CD146 is specifically up-regulated on tumors of neuroectodermal origin and in animal models confers metastatic capacity to human melanoma cells. To identify critical regions regulating MCAM expression in melanomas, 1 kilobase of the MCAM 5' region was analyzed for promoter activity and transcription factor binding in 1 glioma, 1 carcinoma, and 4 melanoma cell lines. The minimal MCAM promoter (106/+22 base pair (bp)) consists of 4 Sp-1 sites, two AP-2 elements, one cAMP responsive element, and the initiator surrounding the transcriptional start site. Analysis of mutated constructs indicated that the cAMP-responsive element is a major transcriptional activator in the majority of cell lines. Site-directed mutagenesis revealed that, in AP-2 expressing cells, the AP-2 site within the core promoter (23 bp) has an inhibitory influence on MCAM expression while the AP-2 sites at 131 and 302 bp are activating. Functional AP-2 was observed in both MCAM positive and MCAM negative melanoma cell lines indicating that expression of MCAM does not require loss of this transcription factor. Furthermore, all MCAM constructs were strongly expressed in MCAM negative as well as MCAM positive cells, indicating that the expression of this gene is not controlled solely by the presence of transactivating factors binding to the investigated region.

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