Golob, Michaela and Buettner, Reinhard and Bosserhoff, Anja-Katrin (2000) Characterization of a transcription factor binding site, specifically activating MIA transcription in melanoma. JOURNAL OF INVESTIGATIVE DERMATOLOGY, 115 (1). pp. 42-47. ISSN 0022-202X
Full text not available from this repository. (Request a copy)Abstract
We have previously isolated the protein MIA, which is secreted from melanoma cells, and identified highly restricted expression patterns in melanocytic tumors. Preliminary studies of the human MIA gene provided evidence that the promoter is specifically activated in melanoma cells but is silent in nonmelanocytic cells and benign melanocytes. In this study we aimed to identify cis-regulatory promoter elements that mediate promoter activation during malignant transformation of melanocytes. We therefore subcloned 1.4 kb of the murine MIA promoter and a series of 5' deletion constructs into a luciferase reporter plasmid and identified the most active cis-regulatory element between nucleic acids -230 and -130. Cloning oligomeric fragments of this promoter region in front of a minimal TK promoter revealed a 30 bp enhancer element mediating expression of the reporter gene in melanoma cells but not in melanocytes or nonmelanocytic cells. Gel mobility shift assays and southwestern blots led to the identification of specific DNA-protein complexes in melanoma cells. Fine mutational analysis of the cis-regulatory promoter element showed that two critical nucleic acid residues are essential for both transcriptional activity and formation of the band shift complexes. By an initial small-scale affinity purification we were able to isolate a protein approximately 32 kDa in size from melanoma cells, which we refer to as MATF (''melanoma-associated transcription factor''). Our study identified for the first time MATF, a transcription factor that is upregulated or activated during malignant transformation of melanocytes.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | CELL-SPECIFIC EXPRESSION; SENSITIVE PROTEIN GENE; MOUSE TYROSINASE GENE; RETINOIC ACID; INHIBITING ACTIVITY; MALIGNANT-MELANOMA; MICROPHTHALMIA; PROMOTER; IDENTIFICATION; CARTILAGE; cell-specific expression; melanoma; MIA; transcription factor |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Pathologie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 05 Apr 2022 08:49 |
| Last Modified: | 05 Apr 2022 08:49 |
| URI: | https://pred.uni-regensburg.de/id/eprint/42357 |
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