Title | Activation of the TSG-6 gene by NF-IL6 requires two adjacent NF-IL6 binding sites. |
Publication Type | Journal Article |
Year of Publication | 1995 |
Authors | Klampfer L, Chen-Kiang S, Vilcek J |
Journal | J Biol Chem |
Volume | 270 |
Issue | 8 |
Pagination | 3677-82 |
Date Published | 1995 Feb 24 |
ISSN | 0021-9258 |
Keywords | Base Sequence, Binding Sites, CCAAT-Enhancer-Binding Proteins, Cell Adhesion Molecules, Cells, Cultured, DNA-Binding Proteins, HeLa Cells, Humans, Molecular Sequence Data, Mutation, Nuclear Proteins, Oligodeoxyribonucleotides, Promoter Regions, Genetic, Protein Binding, Transcription Factors |
Abstract | Tumor necrosis factor (TNF)-stimulated gene 6 (TSG-6) encodes a protein expressed during inflammation. We have previously shown that transcription factors of the NF-IL6 and AP-1 families cooperatively modulate activation of the TSG-6 gene by TNF or interleukin 1 (IL-1) through a promoter region that contains an NF-IL6 site (-106 to -114) and an AP-1 element (-126 to -119). In this study we report the identification of an additional NF-IL6 site (NF-IL6*) located at positions -92 to -83. Footprinting and electrophoretic mobility shift assay suggested that NF-IL6 binds with higher affinity to the newly identified NF-IL6* site than to the earlier identified promoter-distal NF-IL6 site and that the two sites cooperate in binding NF-IL6. TNF and IL-1 stimulate specific binding of nuclear proteins to the NF-IL6* site more efficiently than to the promoter-distal NF-IL6 site. Moreover, a mutation in the NF-IL6* site abolished transactivation of the TSG-6 promoter by NF-IL6 despite the presence of the intact promoter-distal NF-IL6 site. A mutation in the promoter-distal NF-IL6 site also greatly decreased activation of the TSG-6 promoter by NF-IL6. We conclude that the two NF-IL6 sites are functionally interdependent in the activation of the TSG-6 gene. |
DOI | 10.1074/jbc.270.8.3677 |
Alternate Journal | J Biol Chem |
PubMed ID | 7876106 |
Grant List | R35-CA49731 / CA / NCI NIH HHS / United States |
Related Faculty:
Selina Chen-Kiang, Ph.D.