Abstract
Overexpression of ras protooncogenes has been implicated in cancer development. We therefore initiated a study of the human N-ras promoter to determine the regions that control N-ras expression and their potential for interaction with DNA-binding proteins. N-ras CAT constructs were stably integrated into K562 cells by electric field-mediated gene transfer. A significant proportion of promoter activity was found to lie within a 438bp fragment comprising an untranslated exon (exon -1) with adjacent 5′ sequence and a small CpG island. A 107bp fragment at the 5′ end of exon -1 was essential for promoter activity, while a 44bp deletion from within this region decreased promoter activity by two thirds. Unlike the human H-ras promoter, the human N-ras promoter did not exhibit bidirectional activity. DNase footprinting of the 438bp fragment revealed seven protected regions, many of which contain sequences homologous to known DNA binding protein sites (MLTF/myc, CREB/ATF, AP-1, AP-2, myb, and E4TF1). Using purified MLTF and appropriate competitors in gel shift and DNase footprinting assays, we demonstrated binding of MLTF to the MLTF consensus sequence within exon -1.
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© 1991 Plenum Press, New York
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Thorn, J.T., Todd, A.V., Warrilow, D., Watt, F., Molloy, P.L., Iland, H.J. (1991). Characterization of the Human N-ras Promoter Region. In: Spandidos, D.A. (eds) The Superfamily of ras-Related Genes. NATO ASI Series, vol 220. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-6018-6_11
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DOI: https://doi.org/10.1007/978-1-4684-6018-6_11
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