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The Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104 [H. H., A. H. R., K. H., M. I., S. W., R. P. R., Y. T., C. M. C., H. K.]; Department of Microbiology, Kobe University School of Medicine, Kobe 650, Japan [H. H.]; and Department of Cell Biology and Anatomy, Cornell University Medical College, New York, New York 10021 [M. V. C.]
The melanoma-associated antigen ME491 is expressed strongly during the early stages of tumor progression. The ME491 gene was molecularly cloned by means of DNA-mediated gene transfer followed by screening a 
genomic library with human repetitive Alu sequences as a probe. The cloned DNA, after transfection into mouse L-cells, generated a protein with characteristics that were indistinguishable in Western blot analysis from the ME491 antigen expressed by human melanoma cells. Repeat-free subfragments of the cloned DNA were used for further studies. By Northern blot analysis, the subfragments detected a single 1.2-kilobase mRNA in the transformants and various human melanoma cell lines. ME491 complementary DNA clones were then obtained by probing a melanoma complementary DNA library with the genomic subfragments. Nucleotide sequence analysis of the cloned complementary DNA indicated that the ME491 antigen consists of 237 amino acids (Mr 25,475) with four transmembrane regions and three putative N-glycosylation sites. No significant structural homology was observed with other proteins thus far reported. We observed that the amounts of mRNA varied greatly with different melanoma cell lines. Southern blot analysis revealed no amplification or rearrangement of the ME491 gene in the human melanoma cell lines tested, including both high and low expressors of this antigen. The ME491 gene has been mapped to chromosome region 12p12
12q13 by somatic cell hybrid analysis and more narrowly localized to 12q1212q14 by in situ hybridization.
1 Supported by NIH Grants NS-21716, CA-25874, CA-10815, and RR-05540 and grants from the National Neurofibromatosis Foundation and the Dysautonomia Foundation.
2 To whom requests for reprints should be addressed, at The Worcester Foundation for Experimental Biology, 222 Maple Ave., Shrewsbury, MA 01545.
Received 2/24/88. Accepted 3/25/88.
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