CLONING AND ANALYSIS OF MART-1 MELAN-A HUMAN-MELANOMA ANTIGEN PROMOTER REGIONS/

The MART-1/Melan-A human melanoma tumor antigen can be recognized by T lymphocytes and appears to be involved in tumor regression. To study the transcriptional regulation of this important gene, the 5' untransl ated (UT) region of the MART-1/Melan-A gene was cloned and sequenced. Human melanoma cell lines were screened for MART-1/Melan-A mRNA expres sion. Primer extension and northern analysis were performed to confirm the mRNA size and start site. Several overlapping fragments of 5'UT w ere isolated from genomic DNA by polymerase chain reaction (PCR) from the previously described sequence for an additional 700 bp upstream. T he fragments isolated (ranging from 838 bp to 160 bp in length) were u sed to drive luciferase reporter gene expression in melanoma and non-m elanoma cell lines. Tissue-specific promoter activity was found in a 2 33-bp fragment of 5' UT with an average index of induction of 35 fold. The 233-bp MART-1/Melan-A promoter does not appear to have cytokine ( IL-2, IL-4, IL-7, GM-CSF, TNF-alpha or IFN-gamma) responsive elements when studied in transient transfection assays. (C) 1997 Elsevier Scien ce B.V.