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Volume 271, Number 37, Issue of September 13, 1996 pp. 22915-22922
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Identification and Characterization of the Cell Type-specific and Developmentally Regulated 7 Integrin Gene Promoter

(Received for publication, March 15, 1996, and in revised form, June 10, 1996)

From the Departments of Stomatology and Anatomy, University of California, San Francisco, California 94143-0512

Expression of 7 is mainly confined to skeletal and cardiac muscle in which it appears to be the major laminin-binding integrin. When myoblasts differentiate to myotubes, 7 mRNA and protein expression is up-regulated. To explore the mechanisms involved in the tissue-specific and developmentally regulated expression of 7, we isolated and characterized a genomic clone containing ~2.8 kilobase pairs (kb) of the 5-flanking region of the murine 7 gene. The 5-flanking region lacks both TATA and CCAAT boxes but contains five putative Sp1 binding sites located in a CpG island. Two transcription start sites, located near an initiator-like sequence, are 176 and 170 base pairs upstream of the translation start site. There are numerous binding sites for developmental and cell type-specific transcription factors, including AP-1, AP-2, GATA, and several AT-rich sites. There are also eight consensus E-boxes that bind the basic helix-loop-helix family of muscle-specific transcription factors. The ~2.8-kb 5-flanking region was an active promoter in C2C12 skeletal myoblasts and exhibited increased expression upon conversion to myotubes but was inactive in HtLM2 cells, a mouse breast carcinoma epithelial cell line that does not express 7. Deletion analysis identified both positive and negative regulatory elements within the ~2.8-kb fragment. In 10T1/2 fibroblasts the ~2.8-kb 7 promoter was trans-activated by the myogenic basic helix-loop-helix proteins myogenin and MyoD but not by MRF4 and myf5. These results suggest that muscle-specific transcription factors play a role in regulating the cell-type expression of the 7 gene during development.

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