Pendergrast P S, Morrison D, Tansey W P, Hernandez N
Howard Hughes Medical Institute, Cold Spring Harbor, New York 11724, USA.
J Virol. 1996 Aug;70(8):5025-34. doi: 10.1128/JVI.70.8.5025-5034.1996.
The human immunodeficiency virus type 1 promoter generates two types of RNA molecules, full-length transcripts and short transcripts. Synthesis of the short transcripts depends on the inducer of short transcripts (IST), an element located downstream of the start site. In the presence of the viral activator Tat, the synthesis of full-length transcripts is up-regulated while that of short transcripts is down-regulated. Full-length and short transcripts are probably generated by different types of transcription complexes. The first is IST independent, capable of efficient elongation, and up-regulated by Tat. The second is IST dependent, incapable of efficient elongation, and down-regulated by Tat. We have used an in vivo assay to assess the role of TBP in human immunodeficiency virus type I transcription and to test the effect of mutations in TBP on synthesis of full-length and short transcripts. We find that TBP bound to the TATA box is required for the synthesis of short and full-length transcripts as well as for Tat activation and that both yeast TBP and the carboxy-terminal domain of human TBP can replace full-length human TBP for these processes. Mutations in TBP affect the synthesis of short and full-length transcripts as well as Tat activation similarly, and these effects correlate with the previously described effects of these mutations on binding of TBP to the TBP-associated factor TAFII250 in vitro. Together, these results suggest that if short and full-length transcripts are generated by variant transcription complexes, these complexes use TBP similarly, probably as part of the TFIID complex.
1型人类免疫缺陷病毒启动子产生两种类型的RNA分子,即全长转录本和短转录本。短转录本的合成依赖于短转录本诱导因子(IST),这是一个位于起始位点下游的元件。在病毒激活因子Tat存在的情况下,全长转录本的合成上调,而短转录本的合成下调。全长转录本和短转录本可能由不同类型的转录复合物产生。第一种不依赖IST,能够高效延伸,并被Tat上调。第二种依赖IST,不能高效延伸,并被Tat下调。我们利用体内试验来评估TBP在1型人类免疫缺陷病毒转录中的作用,并测试TBP突变对全长和短转录本合成的影响。我们发现,结合到TATA框的TBP对于短转录本和全长转录本的合成以及Tat激活都是必需的,并且酵母TBP和人类TBP的羧基末端结构域都可以替代全长人类TBP进行这些过程。TBP中的突变对短转录本和全长转录本的合成以及Tat激活有类似的影响,并且这些影响与先前描述的这些突变在体外对TBP与TBP相关因子TAFII250结合的影响相关。总之,这些结果表明,如果短转录本和全长转录本由变体转录复合物产生,那么这些复合物对TBP的使用方式类似,可能是作为TFIID复合物的一部分。