A serum response element and a binding site for NF-Y mediate the serum response of the human thrombospondin 1 gene
| Autor: | Paul Framson, Paul Bornstein |
|---|---|
| Rok vydání: | 1993 |
| Předmět: |
Blood Platelets
endocrine system Serum Response Factor Mutant Molecular Sequence Data Oligonucleotides Platelet Membrane Glycoproteins Biology Regulatory Sequences Nucleic Acid Biochemistry Mice Sequence Homology Nucleic Acid Consensus sequence Gene family Animals Humans Binding site Promoter Regions Genetic Molecular Biology Transcription factor Thrombospondin Binding Sites Base Sequence virus diseases Nuclear Proteins Cell Biology 3T3 Cells Serum Response Element Molecular biology DNA-Binding Proteins Blood Gene Expression Regulation Regulatory sequence CCAAT-Enhancer-Binding Proteins Thrombospondins Transcription Factors |
| Zdroj: | The Journal of biological chemistry. 268(7) |
| ISSN: | 0021-9258 |
| Popis: | The expression of thrombospondin 1 (TSP 1), a member of the TSP gene family, is rapidly induced by growth factors. We tested the ability of human TSP 1-chloramphenicol acetyltransferase constructs to respond to serum in stably transfected NIH-3T3 cells. Two transcriptional elements in the TSP 1 promoter, a distal element at -1280 and a proximal element at -65, were required for the response of the human TSP 1 gene to serum. The distal element contains the 5'-CC(A + T)6GG-3' consensus sequence characteristic of a serum-response element (SRE). Deletions or mutations in this element reduced the serum response of the TSP 1 gene by 80-90%. In gel-shift assays, the -1280 element and the c-fos SRE cross-competed, whereas their functional and binding mutants did not. The proximal element contains the sequence 5'-GGCCAATGGG-3', which closely resembles the consensus binding motif for the CCAAT-binding factor NF-Y (CBF, CP1, alpha CP1). Deletions or mutations in this element also reduced the serum response by 80-90%. Methylation interference analysis of the -65 region identified a pattern of contacts with nuclear factors resembling that for NF-Y, and an NF-Y-binding site and the proximal TSP 1 element cross-competed in gel-shift assays, whereas their binding mutants did not. Finally, an abbreviated TSP 1 promoter/5'-flank, containing the SRE- and NF-Y-binding sites, mediated a serum response that was close in magnitude to that of the parent promoter. We conclude that the serum response of the human TSP 1 gene requires the coordinated function of an SRE- and NF-Y-binding site. |
| Databáze: | OpenAIRE |
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