Transactivation of heterologous promoters by HIV-1 tat.
To determine whether HIV-1 tat can transactivate a heterologous promoter lacking HIV sequences other than the TAR element, TAR was placed downstream of the chicken beta-actin promoter. Tat increased expression directed by the actin-TAR promoter to a degree equal to tat induction of the HIV-1 LTR. Optimal transactivation was observed when TAR was positioned downstream of the actin promoter such that the expected cap site of transcripts from this promoter would be the same as in transcripts directed by the HIV-1 LTR. Tat was able to transactivate, though to a lesser extent, a promoter consisting solely of a TATA element fused to TAR. Thus, tat induction does not require HIV-1 LTR promoter sequences other than TAR. Tat, when fused to the DNA binding domain of BPV-1 E2, was able to transactivate a truncated SV40 promoter containing upstream E2 binding sites, indicating that tat may be capable of transactivation when directed by a DNA binding protein to an upstream site in a heterologous promoter lacking all HIV sequences. Substitution of Ala for Lys at position 41 of tat in the tat-E2 fusion, a mutation which dramatically decreases tat transactivation of the HIV-1 LTR, eliminated this transactivation.
Han
P
P
Biogen, Inc., Fourteen Cambridge Center, MA 02142.
Brown
R
R
Barsoum
J
J
D016475:3T3 Cells; D000199:Actins / Q000235:genetics; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D052688:Bovine papillomavirus 1 / Q000235:genetics; D002645:Chickens; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics; D004742:Enhancer Elements, Genetic / Q000235:genetics; D015967:Gene Expression Regulation, Viral / Q000235:genetics*; D015696:Gene Products, tat / Q000235:genetics*; D016325:HIV Long Terminal Repeat / Q000235:genetics; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics; D015534:Trans-Activators / Q000235:genetics; D015533:Transcriptional Activation / Q000235:genetics*; D014162:Transfection / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics; D054322:tat Gene Products, Human Immunodeficiency Virus
D000199:Actins; D004268:DNA-Binding Proteins; C051550:E2 protein, Bovine papillomavirus; D015696:Gene Products, tat; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D014764:Viral Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus; D015500:Chloramphenicol O-Acetyltransferase