A novel protein interacts with the major transforming growth factor-beta responsive element in the plasminogen activator inhibitor type-1 gene.
Multiple transforming growth factor-beta (TGF-beta) responsive elements have been identified within the 5'-flanking region of the plasminogen activator inhibitor type-1 (PAI-1) gene. This study was designed to characterize the major TGF-beta responsive element (-804 to -546). DNA footprint assays showed that the region of protein contact (-726 to -703) did not include consensus sequences for any known transacting factors. The results of UV cross-linking and Southwestern blot experiments showed that a protein of M(r) 100,000 specifically binds to the TGF-beta responsive element and that this protein undergoes post-transcriptional activation within 5 min after stimulation of Hep G2 cells by TGF-beta resulting in a marked increase in affinity for the target DNA sequence. These results show that stimulation of Hep G2 cells with TGF-beta increases the affinity of a novel 100-kDa protein for the major TGF-beta responsive element within the PAI-1 gene.
Sandler
M A
MA
Cardiovascular Division, Washington University School of Medicine, St. Louis, Missouri 63110.
Zhang
J N
JN
Westerhausen
D R
DR
Billadello
J J
JJ
D001483:Base Sequence; D002478:Cells, Cultured; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005786:Gene Expression Regulation; D006801:Humans; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics*; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D014466:Ultraviolet Rays
D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D017395:Plasminogen Activator Inhibitor 1; D016212:Transforming Growth Factor beta