Characterization of the Human N-ras Promoter Region

  • J. T. Thorn
  • A. V. Todd
  • D. Warrilow
  • F. Watt
  • P. L. Molloy
  • H. J. Iland
Part of the NATO ASI Series book series (NSSA, volume 220)


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.


K562 Cell Negative Regulatory Element Untranslated Exon Major Late Promoter Human Fibrosarcoma Cell Line HT1080 


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Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • J. T. Thorn
    • 1
  • A. V. Todd
    • 1
  • D. Warrilow
    • 1
  • F. Watt
    • 2
  • P. L. Molloy
    • 2
  • H. J. Iland
    • 1
  1. 1.The Kanematsu LaboratoriesRoyal Prince Alfred HospitalCamperdownAustralia
  2. 2.CSIRO Division of Biomolecular EngineeringNorth RydeAustralia

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