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A novel SP-1 site in the human interleukin-1β promoter confers preferential transcriptional activity in keratinocytes

✍ Scribed by Matthias Husmann; Petra Jehnichen; Bernhard Jahn; Dominik Schlosshan; Eric Romahn; Jürgen Marx

Publisher
John Wiley and Sons
Year
1996
Tongue
English
Weight
742 KB
Volume
26
Category
Article
ISSN
0014-2980

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✦ Synopsis

A novel SP-1 site in the human interleukin-1p promoter confers preferential transcriptional activity in keratinocytes

To investigate the mechanisms of transcriptional activation of interleukin-1P (IL-1p) in non-monocytic cells, we constructed a series of reporter plasmids with the bacterial chloramphenicol acetyltransferase gene linked to various parts of the human IL-1p promoter and performed transient transfection experiments. We identified a promoter segment that activates transcription most efficiently in keratinocytes. Electrophoretic mobility shift assays (EMSA) with a 43-mer oligonucleotide derived from the functionally identified cis-acting element revealed specific complexes. By competition analysis with transcription factor consensus sequence oligonucleotides and by immunosupershift, transcription factor SP-1 or a closely related protein was shown to bind to this regulatory element. The closest match to the known SP-1 consensus sequence within the respective region is a TCCCCTCCCCT motif. Mutation of this motif almost completely, and specifically, abolished the binding of two low-mobility complexes and led to a 95 % decrease of constitutive transcriptional activation of a reporter construct IL-1p (-170/+ 108). Likewise, activation of this reporter construct by tumor necrosis factor-a depended on the SP-1 site. These observations suggest that a so-far-unrecognized SP-1 site in the human IL-lp promoter may participate in the transcriptional regulation of this gene in keratinocytes.

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