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  • Original Paper
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The EGF/TGFα response element within the TGFα promoter consists of a multi-complex regulatory element

Oncogene volume 18, pages 5923–5935 (1999)Cite this article

Abstract

Autocrine TGFα is an important growth effector in the transformed phenotype. Growth stimulation of some colon cancer cells as well as other types of cancer cells is effected by activation of the epidermal growth factor receptor. Importantly, this receptor activation leads to further stimulation of TGFα transcription and increased peptide synthesis. However, the molecular mechanism by which TGFα transcription is activated is poorly understood. In this paper, we describe the localization of a cis-sequence within the TGFα promoter which mediates this stimulation. This region contains parallel cis-acting elements which interact to regulate both basal and EGF-induced TGFα expression. The well differentiated colon carcinoma cell line designated FET was employed in these studies. It produces autocrine TGFα but requires exogenous EGF in the medium for optimal growth. Addition of EGF to FET cells maintained in the absence of EGF resulted in a 2 – 3-fold increase of both TGF promoter activity and endogenous TGFα mRNA at 4 h. This addition of EGF also stimulated protein synthesis. The use of deletion constructs of the TGFα promoter in chimeras with chloramphenicol acetyl transferase localized EGF-responsiveness to between −247 and −201 within the TGFα promoter. A 25 bp sequence within this region conferred EGF-responsiveness to heterologous promoter constructs. Further use of deletion/mutation chimeric constructs revealed the presence of at least two interacting cis-elements, one binding a repressor activity and the other, an activator. Gel shift studies indicate the presence of distinct complexes representing activator and repressor binding, which are positively modulated by EGF. The type and amount of complexes formed by these proteins interact to regulate both the basal activity and EGF-responsiveness of the TGFα promoter. The interaction of an activator protein with an EGF-responsive repressor may serve to regulate the level of this progression-associated, transforming protein within tight limits.

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Abbreviations

TGFα:

transforming growth factor α

EGF:

epidermal growth factor

EGFr:

epidermal growth factor receptor

bp:

base pair(s)

CAT:

chloramphenicol acetyltransferase

tk:

thymidine kinase

cAMP:

cyclic adenosine monophosphate

Neo:

Neomycn

PCR:

polymerase chain reaction

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Acknowledgements

This work was supported by National Institutes of Health Grants CA34432 and CA54807. Work performed in partial fulfillment of the requirements for the PhD degree for R Awwad. We thank Suzanne Payne, Jenny Zak and Lorraine Gilmore for preparation of the manuscript and Barbara Young for excellent technical assistance.

Author information

Author notes

  1. Lisa E Humphrey

    Present address: Department of Surgery, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, Texas, TX 78284, USA

  2. Rana Awwad, Lisa E Humphrey, Wenhui Li and Gillian M Howell: R Awwad, LE Humphrey and GM Howell contributed equally to this work

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Medical College of Ohio, PO Box 10008, Toledo, 43699-0008, Ohio, OH, USA

    Rana Awwad, Lisa E Humphrey, Baskar Periyasamy, William Scovell Jr, Wenhui Li, Michael G Brattain & Gillian M Howell

  2. Department of Molecular Genetics, Oncology Drug Discovery, Bristol Myers Squibb, PO Box 4000, Princeton, 08543-4000, New Jersey, NJ, USA

    Rana Awwad, Mark Lynch & Joan Carboni

  3. Box 123, Pfizer Central Research, Eastern Point Road, Groton, Connecticut, 06340, CT, USA

    Kevin Coleman

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Awwad, R., Humphrey, L., Periyasamy, B. et al. The EGF/TGFα response element within the TGFα promoter consists of a multi-complex regulatory element. Oncogene 18, 5923–5935 (1999). https://doi.org/10.1038/sj.onc.1202982

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