PMID: 9550409Apr 29, 1998Paper

Characterization of the IFN-gamma-responsive element in the 5' flanking region of the C1 inhibitor gene

The Journal of Immunology : Official Journal of the American Association of Immunologists
Kamyar ZahediAlvin E Davis

Abstract

Treatment of a variety of cell lines with IFN-gamma leads to enhanced synthesis and secretion of C1 inhibitor (C1inh). The induction of C1inh synthesis by IFN-gamma is primarily regulated at the transcriptional level, and is controlled by elements in the 5' flanking region and the first intron of the C1inh gene. Hep3B cells transfected with reporter constructs containing truncated segments between -738 and -81 of the 5' flanking region and stimulated with IFN-gamma expressed increased levels of chloramphenicol acetyl transferase. These data as well as the data obtained from studies using constructs with mutated IFN-gamma-activated sequence (GAS) indicate that the most proximal GAS element (GAS 4) that mapped to nucleotides -126 to -118 is responsible for this IFN-gamma responsiveness. Electrophoretic mobility shift assays using GAS 4 yielded a single band that appeared within 5 min after stimulation with IFN-gamma. In competition experiments, both GAS 4 and consensus GAS probes, but not a mutated GAS probe, competed for the GAS binding protein present in Hep3B and U-937 cell extracts. The identity of the GAS binding protein was confirmed using anti-STAT-1alpha Abs in supershift assays. The results indicate that STAT-1alpha bind...Continue Reading

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