Integrin _v Promoter Activity in Keratinocytes

ISSN： 0022-4804

Source： Journal of Surgical Research, Vol.76, Iss.2, 1998-05, pp. : 185-191

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Abstract

Background. During reepithelialization keratinocytes show increased expression of the integrin subunit _v. We have investigated the promoter region of the _v integrin subunit to learn more about its regulation.Methods. The promoter region of the human integrin _v gene was cloned into a luciferase reporter vector. Deletional mutants were created using PCR. Computerized sequence analysis was performed using the Wisconsin Package. Gel-shift analysis was performed using keratinocyte nuclear extracts and oligonucleotides spanning the regions of interest.Results. Deletion from -522 bp to -235 resulted in no discernible effect on promoter activity. In contrast deletion of the next 22 bp, which included a putative ets binding site, reduced activity by approximately half. Further deletion to -139 bp essentially abolished promoter activity. Computer searching of this region of the integrin _v promoter revealed two tandemly repeated motifs, TCCTCCTCC, that had previously been implicated in the function of the epidermal growth factor receptor (EGFR) promoter. Comparison of the _v integrin promoter to the EGFR promoter revealed an area of high homology in this region. Gel-shift analysis revealed binding of a single-strand specific DNA binding protein to single-stranded oligos comprising these motifs, but no binding of factors to the double-stranded oligo containing the ets binding site.Conclusions. In keratinocytes _v integrin expression is controlled by a region of the promoter with high homology to the epidermal growth factor receptor promoter. This region binds single-strand specific DNA binding proteins that are likely to be important in controlling transcription.