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| Content Provider | Journal of Biological Chemistry (JBC) |
|---|---|
| Author | Ruocco, Maria Rosaria Chen, Xueni Ambrosino, Concetta Dragonetti, Emila Liu, Weimin Mallardo, Massimo Falco, Giulia De Palmieri, Camillo Franzoso, Guido Quinto, Ileana Venuta, Salvatore Scala, Giuseppe |
| Abstract | We report the characterization of a CAAT enhancer-binding protein (C/EBP) (NF-IL6) element encompassing the region from −174 to −166 of the U3 long terminal repeat (LTR) region of HIV-1. This C/EBP cis sequence was found to bind to C/EBPβ and C/EBPδ factors in DNA band shift assay. Transfection of NTera-2 cells with a HIV-1-LTR CAT construct (pC15CAT), together with C/EBPβ or C/EBPδ expression plasmids showed that C/EBP proteins strongly activated the HIV-1 promoter. Deletions encompassing the C/EBP-binding site resulted in the enhancement of the LTR activation mediated by C/EBP proteins, suggesting that other sequences located 3′ to −170 were indeed the target for C/EBP factors. This possibility was confirmed by using the pCD54E9CAT plasmid, in which the NF-κB enhancer was inserted 5′ to the HIV-1 LTR TATA box. A NF-κB1(p50) expression plasmid was also utilized to test for functional co-operation between NF-κB and C/EBP factors. We observed that p50·C/EBPβ and p50·C/EBPδ complexes were generated in tested cells and strongly activated the HIV-1 LTR by binding to the NF-κB sequences. The physical association of NF-κB1(p50) with C/EBP factors was assayed by direct interaction of in vitro translated p50 proteins with C/EBPβ or C/EBPδ produced as glutathione S-transferase fusion proteins. Moreover, p50·C/EBPβ complexes were observed in vivo by using DNA affinity studies with biotinylated NF-κB oligonucleotides. By using mutant forms of p50 or C/EBPβ proteins we found that the transactivation of HIV-1 LTR by p50·C/EBPβ complexes required the DNA-binding domain of p50 and the transcription activation domain of C/EBPβ. |
| Related Links | http://www.jbc.org/content/271/37/22479.abstract |
| Ending Page | 22486 |
| Starting Page | 22479 |
| Page Count | 8 |
| File Format | HTM / HTML PDF |
| ISSN | 00219258 |
| Journal | Journal of Biological Chemistry (JBC) |
| Issue Number | 37 |
| Volume Number | 271 |
| DOI | 10.1074/jbc.271.37.22479 |
| e-ISSN | 1083351X |
| Language | English |
| Publisher | American Society for Biochemistry and Molecular Biology |
| Publisher Date | 1996-09-13 |
| Access Restriction | Open |
| Subject Keyword | Nucleic Acids, Protein Synthesis, and Molecular Genetics |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology |
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