Regulation of HIV-1 Long Terminal Repeats by Interaction of C/EBP(NF-IL6) and NF-κB/Rel Transcription Factors*

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.

• Publication year

  1996

• Venue

  Journal of Biological Chemistry

• Publication date

  1996-09-13

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1074/JBC.271.37.22479PMID 8798413
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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