Olsen H S, Rosen C A
Department of Gene Regulation, Roche Institute of Molecular Biology, Nutley, New Jersey 07110-1199.
J Virol. 1992 Sep;66(9):5594-7. doi: 10.1128/JVI.66.9.5594-5597.1992.
Tat-mediated transcriptional activation of human immunodeficiency virus (HIV) gene expression requires the presence of the cis-acting Tat-responsive element, TAR, and a functional enhancer-promoter element. The ability of Tat to function with heterologous enhancer sequences led us to examine the role of the minimal basal promoter for trans activation. Substitution of HIV TATA sequences (nucleotides -20 to -35) with TATA elements derived from other promoters had little effect on the basal level of transcription or the ability to activate the HIV long terminal repeat upon stimulation through upstream activation sequences. In contrast, minimal alterations within the TATA motif had a profound effect on trans activation, as demonstrated by the 3- to 10-fold reduction in activation following expression of Tat. Our findings suggest that minor changes in the TATA motif affect the composition of the initiation-elongation complex and that the composition of this complex is critical for Tat-dependent activation of gene expression.
由反式激活因子(Tat)介导的人类免疫缺陷病毒(HIV)基因表达的转录激活需要顺式作用的Tat反应元件(TAR)和功能性增强子-启动子元件的存在。Tat与异源增强子序列协同作用的能力促使我们研究最小基础启动子在反式激活中的作用。用源自其他启动子的TATA元件替换HIV的TATA序列(核苷酸-20至-35)对基础转录水平或通过上游激活序列刺激后激活HIV长末端重复序列的能力影响很小。相反,TATA基序内的微小改变对反式激活有深远影响,Tat表达后激活作用降低3至10倍就证明了这一点。我们的研究结果表明,TATA基序中的微小变化会影响起始-延伸复合物的组成,并且该复合物的组成对于Tat依赖性基因表达激活至关重要。